Toxoplasma gondii is an obligate intracellular parasite that causes toxoplasmosis, with approximately one third of the population around the world seropositive. The consumption of contaminated food is the main source of infection. These include meat products with T. gondii tissue cysts, and dairy products with tachyzoites. Recently, contamination has been detected in fresh products with oocysts and marine products. Despite the great health problems that are caused by T. gondii, currently there are no standardized methods for its detection in the food industry. In this review, we analyze the current detection methods, the prevalence of T. gondii in different food products, and the control measures. The main detection methods are bioassays, cell culture, molecular and microscopic techniques, and serological methods, but some of these do not have applicability in the food industry. As a result, emerging techniques are being developed that are aimed at the detection of multiple parasites simultaneously that would make their application more efficient in the industry. Since the prevalence of this parasite is high in many products , it is necessary to standardize detection methods, as well as implement control measures. Toxoplasma gondii. This protozoon of the Apicomplexa phyla presents only felines as the definitive host, being the ones where the parasite can complete its life cycle. However, all warm-blooded animals, including mammals and birds, can act as intermediate hosts  used a recombinant dense granule antigen 7 protein for the detection of parasites in blood samples [T. gondii are necessary [T. gondii infection through food, prevalence of different food products, its detection and control, and future perspectives.However, and despite the great health public problem that it poses, there are currently no specific detection criteria for at costs . The altat costs . Other sat costs ,25. The  samples . In addiecessary , and theT. gondii is a high priority foodborne zoonotic pathogen around the world, it is not systematically controlled [T. gondii in any food matrix [T. gondii DNA in samples, while information on the viability and infectivity can be obtained by in vivo assays  or by in vitro culture techniques. A summary of these methods with sensitivity and type of food product where these methods have been used are shown in Although ntrolled . At presd matrix . Even soT. gondii detection, the cat bioassay works best, followed by the mouse bioassay [T. gondii oocysts, and serum samples may be analyzed to detect specific antibodies against the parasite [This method allows the study of the infectivity of the oocysts and tissue bradyzoites of the parasite. For bioassay . In catsparasite . The catparasite . Howeverparasite .T. gondii oocysts [T. gondii by microscopy or PCR in addition to detect specific antibodies in serum. Immunosuppressive drugs can be administered to mice to increase the sensibility of the bioassay [Therefore, mice are the main animal model to evaluate the infectivity of oocysts. In this technique, 50 to 200 g of tissue are digested with acid pepsin or trypsin and a fraction of the sediment is inoculated in mice, generally intraperitoneally or subcutaneous; although mice can also be infected orally with  oocysts . Typicalbioassay . The sambioassay . These mbioassay , and to bioassay . Bioassabioassay ,59. HoweT. gondii by isolation in cell culture. The test sample is brought into contact with culture of different cell lines. If the parasite is present in the sample and it is viable, the culture cells will become infected, causing the tachyzoites to multiply, which can be observed with an inverted microscope after 3–10 days [T. gondii could be detected [Despite the fact that molecular techniques are very specific and sensitive, they only detect parasite DNA, regardless of whether it is viable. A solution to this problem is the detection of –10 days . Cell cu–10 days . Even so–10 days . Moreovedetected .T. gondii cannot be detected by gross inspection but can be identified by microscopy. In fact, microscopic methods are used for the detection of oocysts in fresh products and shellfish. Parasites are visible using nonspecific stains such as Giemsa or hematoxylin and eosin, but the use of specific stains with fluorescence-conjugated enzymes or antibodies allows them to be differentiated from other structures or apicomplexan parasites and increases the sensitivity [Oocysts, tachyzoites, or tissue cysts of sitivity . The maiThe polymerase chain reaction (PCR) is based on the in vitro amplification of specific DNA sequences. For these sequences, the DNA that is present in the analyzed samples is extracted, and several amplification cycles are carried out. The presence of parasite-specific DNA in the sample is visualized by agarose gel electrophoresis. If the sample contains the target DNA, a specific band is observed in the gel .T. gondii by PCR. The most common are the B1 fragment, which is repeated 35 times in the parasite genome, and a region of 529 bp that is repeated 200–300 times [T. gondii in such a small sample is low. To allow analysis of large samples and to increase the detection sensitivity, methods that are based on artificial digestion, homogenization and isolation on Percoll gradients, and sequence-based magnetic capture have been described [T. gondii and the release of bradyzoites [There are different targets that are available for the detection of 00 times . Howeverescribed . This aldyzoites .T. gondii by qPCR the most widely used fluorophores are SYBR Green and TaqMan probes [T. gondii, in berry fruits [T. gondii, in spinach. This method was found to be more sensitive than traditional qPCR [Quantitative PCR (qPCR) is a variant of conventional PCR, which allows the detection of the parasite DNA concentration in the analyzed sample with elevated sensitivity, precision, and speed than conventional PCR, in addition to not requiring the use of gels. For the detection of n probes ,66. The n probes . The PCRy fruits . The resnal qPCR .T. gondii. For the detection of oocysts in fresh products, an adaptation of the LAMP technique has been developed. This new technique includes a chromatographic detection system with a lateral flow test strip that allows to accelerate the visualization of the results [T. gondii in meat samples. The results suggest that RT-LAMP is a simple and reliable tool to detect meats that are contaminated with T. gondii [Loop-mediated isothermal amplification (LAMP) enables DNA amplification with high sensitivity and specificity, efficiency, and speed . It is a results . In 2013. gondii . LAMP se. gondii .T. gondii is unknown. So, other methods are required to establish whether the detected DNA belongs to viable parasites [These molecular techniques detected the DNA of the parasite, and genotyping is possible with them. However, different available genotyping methodologies have been irregularly applied in different geographic areas and over different matrices . The maiarasites . Until narasites , and itsarasites .T. gondii in animals and humans, but they have been adapted for testing meat and meat juices. Generally, they serve as a first screening to detect seropositive animals, in which later the infection will be confirmed in the tissue samples through a bioassay. However, these methods can also be used to detect infection in meat juice samples, for example [T. gondii in serum or meat juice are indirect hemagglutination antibody (IHA), the latex agglutination test (LAT), indirect fluorescent antibody test (IFAT), modified agglutination test (MAT), Western blot, and ELISA, with MAT, IFAT, and ELISA being the most used and validated methods [T. gondii [Serological methods are indirect methods that are intended to confirm infection with  example . The ser methods . All the methods . The ELI. gondii ,76. The . gondii .T. gondii that improve the efficiency and reproducibility.The traditional methods of detection have limitations and there are no standardized protocols for their application in the food industry. For this reason, new detection methods are being developed for T. gondii in fresh products [T. gondii and other protozoan, with better results for Toxoplasma than other molecular approaches [T. gondii genotypes can provide relevant information for the control of this zoonosis. For this reason, different studies have evaluated detection and genotyping methods. Recently, similar sensitivity and specificity has been observed of the B1 and ROP8 genes for detection, whereas the latter seems more appropriate for genotyping [In fresh products in particular, oocyst detection methods are scarce. In the last years, different authors have been developed methods for their detection. Lalonde and Gajadhar (2016) developed real-time PCR methods for the identification of protozoan oocysts in vegetables and fruits . Slana eproducts . In bivaproaches . The DNAproaches . The detnotyping .T. gondii in food, but it requires further validation before it can be applied to other food samples. Furthermore, since qPCR only allows determining the presence of the parasite, but cannot directly confirm the viability [T. gondii RNA using reverse transcription and subsequent PCR (RT-PCR). This technique uses the enzyme reverse transcriptase to synthetize complementary DNA (cDNA) from the RNA molecules that are present in the sample. Although this technique has a high sensitivity, RNA degrades much faster than DNA and it is more easily contaminated, so this technique must be performed by highly qualified and experienced personnel. It has difficulty detecting tissue cysts, since it needs the parasite to be metabolically active at the time of analysis [One of the most relevant steps for molecular methods detection is the DNA extraction approach, that depends on matrix analyses. However, few studies have been done that are related it. Temesgen et al. (2020) compared two commercially available DNA extraction procedures in berry fruits . On the iability , recentlanalysis . Recentlanalysis  and a neanalysis .T. gondii [T. gondii in serum chicken, and they obtained high sensitivity (90.5%) and specificity (95.4%) [Loreck et al. (2019) developed a protein microarray for the simultaneous detection of IgG antibodies against different zoonotic agents and pathogens that cause disease in pigs, among which is . gondii . This is (95.4%) . The bes (95.4%) .T. gondii, but which is generalized in all foodborne parasites, to a lesser or greater extent. In this context, a novel metabarcoding assay followed by next generation sequencing (NGS) has been developed to simultaneously detect Cryptosporidium spp., Giardia spp., and T. gondii in shellfish [T. gondii . About this, Guggisberg et al. (2020) have fine-tuned a one-way sequential sieving method to identify different stages of parasites in lettuce [Different recent studies are aimed at improving sensitivity, looking for alternatives to bioassays that allow detecting the viability of the parasite, and a possible validation to be able to apply in the food industry. Moreover, the detection of different parasites simultaneously is relevant to the food industry, so the lack of standardized protocols does not only refer to hellfish . The app lettuce . Even soT. gondii to humans [Transmission through food is the main system of transmission of o humans . Tissueso humans , and spoo humans . Next, wT. gondii infections have been reported in all meat production animals around the world, although the prevalence depends on the detection method that is used  has been detected by different methods [T. gondii DNA was 9.2% and 8.1%, respectively [In China, Japan, and the USA, recent studies indicated a 10–20% infection rate of quick-frozen chickens ,130,160. tenella . Not onld organs . But T.  Romania . In the  Romania ,150, whi Romania . In Cana methods . The com methods . In thisectively ,137.T. gondii and a reservoir of the parasite. Wild ruminants have been analyzed in different European countries and the studies showed a high seroprevalence in roe deer (39.6%), fallow deer (37.1%), red deer (16.6%), Southern chamois (14%), mouflon (11.5%), Iberian wild goat (7.8%), Barbary sheep (5.6%) [T. gondii around the world, with a seroprevalence of 76.9% in Brazil [T. gondii in meat for human consumption is high in Asia, Europe, and the USA, with its detection and control before sale being of vital importance. In the last years, several studies have indicated that wildlife can be a source of infection by p (5.6%) , and othp (5.6%) ,147. Frop (5.6%) . Wild bon Brazil , around n Brazil ,145, 14%n Brazil , 15% in n Brazil , and 9% n Brazil . Crotta n Brazil . From thT. gondii in milk samples from sheep, goat, camels, and donkeys [T. gondii antibodies in the goat serum with a prevalence of T. gondii DNA in milk samples [Tachyzoites can be shed in the milk of acutely infected animals, so both raw milk and raw dairy products can pose a risk of infection for consumers  Table 3Table 3.  donkeys ,169,170, donkeys . However donkeys , and wer donkeys . Other s samples .T. gondii through cow’s milk has not been detected [T. gondii DNA does not allow the viability of the parasite to be determined. For this reason, other techniques have been used to determine the viability of parasites in milk and dairy products, including the viability assay in cell culture, where the cytopathic effect of tachyzoites on Hep-2 cells is measured, including mouse and cat bioassays [T. gondii in the milk and cheese of goats, demonstrating that fresh milk and cheese are a source of transmission, so the protozoan survives cold-enzyme treatment [T. gondii antibodies in goat milk samples. The study showed that this technique in milk samples could easily be applied to detect the seroprevalence of T. gondii, although it does not allow the detection of tachyzoites [Although the transmission of detected , tachyzodetected , which cdetected , or the detected . Howeverioassays . Mouse areatment . The ELIhyzoites .T. gondii oocysts from cat feces or contaminated water, and act as a source of infection in humans. Oocyst detection in environmental and food samples is difficult due to complications in separating and concentrating oocysts from complex matrices, such as raw vegetables, so there is a lack of optimized laboratory methods for its detection [T. gondii in water, soil, and food samples , based on methods that are used for other protozoa [T. gondii in experimental contaminated baby ready-to-eat lettuces. The detection limit of this method was approximately 25 oocysts per 50 g of lettuce leaves [T. gondii, detecting two to nine oocysts per gram of spinach [Fresh products can become contaminated with etection . Howeverprotozoa . Hohweyeprotozoa . In addie leaves . Recentl spinach .T. gondii DNA in fruits and vegetables was in 2012 [T. gondii has been observed in different vegetables around the word, including lettuce, chicory, rocket, parsley, spinach, pack choi, cabbage, rape, asparagus, endive, Chinese chives, carrots, cucumbers, strawberries, and radish [The first detection of  in 2012 . Nowaday in 2012 ,179,180. in 2012 . Contamid radish   [In Poland, vegetables from shops and home gardens presented a contamination rate by  of 9.7% . In Chin of 9.7% , whereas of 9.7% . In Ital produce  or it wa produce . The pregetable) . Also wigetable) .T. gondii oocysts. Mollusks such as clams, mussels, oysters, and scallops, filter-feed and trap phytoplankton in the gills. This filter feeding process can also concentrate waterborne pathogens within their tissues, including oocysts, which can survive for long periods of time in both fresh- and salt-water [T. gondii [T. gondii [Aquatic environments can be contaminated with wastewater carrying lt-water . For detlt-water ,190. Var. gondii . In addi. gondii . Serolog. gondii , rather T. gondii infection. The consumption of raw mollusks is considered a risk factor for T. gondii with elevated temperatures and precipitations [T. gondii in fish has been investigated. There is still controversy about possible parasitic infection in cold-blooded hosts. Some studies support that these animals can act as mechanical vectors, such as mollusks, containing oocysts in their digestive system [T gondii DNA has been found in different fish species of local fish markets [Different studies showed the prevalence of infection in Mediterranean bivalves of 6.6% to 9.4% in countries such as Turkey and Italy ,190,198.itations . Similaritations . Recentle system . In fact markets  and mari markets .T. gondii infection must be done at several levels. First, certain risk factors increase the prevalence of the parasite in farm animals. Hygienic management practices and correct management which involves keeping cats away from crops and gardens and animal feed, are essential to control this pathogen in farms [T. gondii infection in humans. In fact, pasteurization of milk and milk products is also an important control measure. Undoubtedly, stopping consuming these types of products could considerably reduce the prevalence of infection in humans. On the other hand, as occurs in meat products, adequate hygienic and sanitary conditions on farms would lead to this reduction. In fresh products and vegetables, the most common mechanism of contamination is irrigation with water that is contaminated by oocysts, so sanitary control measures in irrigation water would be interesting. Furthermore, washing fresh produce after harvest and before consumption is an important control measure, since the chemical disinfectants are not effective [The control of in farms . Temperain farms ,202. Thein farms . In the in farms . Neverthin farms ,112. Conffective .T. gondii in food production is essential. However, control measures during food inspection are not applied [T. gondii can be eliminated from meat in 5–6 min at 49 °C, in 44 s at 55 °C, or in 6 s at 61 °C [T. gondii during cooking and low temperature storage and concluded that the parasite was not found when the internal temperature reached 64 °C and below −18 °C [The control of  applied . Current applied ,203. Rel applied . In factat 61 °C . Differeat 61 °C . Rani anw −18 °C .T. gondii. To inactivate isolate tissue cysts, a minimum of three days is required at −20 ºC [T. gondii in food for thermal and non-thermal methods has been extensively analyzed in the review that was published by Mirza et al. (2018) [However, these elevated temperatures are not applicable to all food matrices. This is the case of vegetables and fresh products . Regardit −20 ºC . In addit −20 ºC ,154,210,t −20 ºC ,212, andt −20 ºC ,75. The . (2018) .T. gondii in food products has been realized traditionally with high temperatures  and when cured and salted [T. gondii in food. High pressure processing (HHP) is a novel method for liquid and solid food products where pressures of 340–550 MPa during 1 min can inactive cysts of the parasite [T. gondii cysts in meat [The inactivation of d salted ,214, wheparasite . The sec in meat . HoweverT. gondii infection is carried out at home, setting recommendations on food consumption in the groups that are most vulnerable to the parasite. This situation occurs because there are no regulations governing control measures against T. gondii in the food industry. Inactivation methods have yet to be optimized and validated to be applied against this parasite is a systematic way. More, different prevention measures could also be applied to its control. In farms, biosecurity and control will be factors of great relevance for infection control. Other measures such as restricting the access of cats to crops, gardens, and livestock feed, or the development of a vaccine that is aimed at cats to prevent the active release of oocysts could be used. Prevention measures could also be implemented at the livestock farm, such as the vaccination of cattle. Today, a vaccine against T. gondii is available for sheep, which prevents the spread of parasites to the placenta, and is used for the prevention of abortions in this species [Currently, most of the control of  species . This va species . This meT. gondii and deriving its use for preheated or frozen meat products since, as we have seen, these methods are effective for the inactivation, as well as marking negative products as free from T. gondii [It would be useful to carry out a follow-up program at slaughter, detecting the meat that is positive for . gondii . DetectiT. gondii is the food parasite with the greatest epidemiological relevance, which is distributed worldwide, and with a complex life cycle that makes its detection very difficult. The main foods that are involved in the transmission of this parasite are meat and fresh products (vegetables and fruits) products through tissue cysts, mollusks, and fish, as well as through oocysts, and milk and dairy products through tachyzoites. Currently, the main detection methods are bioassays, in vitro culture, molecular methods (PCR and LAMP), and microscopy as direct methods, and serological techniques as MAT, IFAT, and ELISA as indirect methods. Due to the limitations of these methods, the emerging detection methods are aimed at developing methods with greater sensitivity and reproducibility, and generally, are aimed at the detection of several parasites simultaneously, which would increase their efficiency and facilitate their application in the food industry. Control methods include thermal methods such as heat, cooking, and freezing, as well as non-thermal methods such as HPP, IR, curing, or salting. Most of the control of T. gondii is carried out at home since there are no microbiological criteria for this parasite in the food industry and, therefore, it is not mandatory to comply with control measures. In the future, new detection methods should be validated to optimize the control of infection in food and apply them in the food industry.


Trichophyton (T.) indotineae is a newly identified dermatophyte species that has been found in a near-epidemic form on the Indian subcontinent. There is evidence of its spread from the Indian subcontinent to a number of countries worldwide. The fungus is identical to genotype VIII within the T. mentagrophytes/T. interdigitale species complex, which was described in 2019 by sequencing the Internal Transcribed Spacer (ITS) region of ribosomal DNA of the dermatophyte. More than 10 ITS genotypes of T. interdigitale and T. mentagrophytes can now be identified. T. indotineae causes inflammatory and itchy, often widespread, dermatophytosis affecting the groins, gluteal region, trunk, and face. Patients of all ages and genders are affected. The new species has largely displaced other previously prevalent dermatophytes on the Indian subcontinent. T. indotineae has become a problematic dermatophyte due to its predominantly in vitro genetic resistance to terbinafine owing to point mutations of the squalene epoxidase gene. It also displays in vivo resistance to terbinafine. The most efficacious drug currently available for this terbinafine-resistant dermatophytoses, based on sound evidence, is itraconazole. Trichophyton (T.) rubrum, the erstwhile predominant dermatophyte not only responsible for tinea pedis and onychomycosis, but also for dermatophytosis involving the whole body, worldwide—including India over the past few decades  in Bethesda, Maryland, USA, which consistently classified all strains within the species complex T. mentagrophytes/T. interdigitale as T. interdigitale until 2016 [T. mentagrophytes (zoophilic) and T. interdigitale (anthropophilic) [The f et al.  can be sof Delhi , in whictil 2016 . It was ophilic) . It has ophilic) . T. mentagrophytes genotype VIII has also recently changed [T. mentagrophytes is now classified as T. indotineae [T. mentagrophytes/T. interdigitale, are also referred to as cryptic species or molecular siblings. It has been suggested that it makes sense to describe such genotypes as independent species only if they have significant morphological differences that have clinical significance . Since that is not the case for most genotypes, it has been suggested to designate these as mere “clonal offshoots” [T. indotineae is morphologically  [The current designation of  changed ,34. Accodotineae   . T. indotineae is mainly transmitted from person to person. Spread of T. indotineae infection as a family case was documented in Iran [T. indotineae in Germany was documented in a tinea-corporis-affected baby from Bahrain and his multiple family members, a German woman and her husband from Saudi Arabia, and from a Libyan to his female partner and their child. According to current knowledge,  in Iran . None of in Iran . TransmiT. indotineae has clearly adapted excellently to the milieu on the human epidermis and is also easily transmitted to other persons directly by physical contact. However, an indirect transmission is also seen via inanimate surfaces in the living environment, such as bathrooms and lavatories, and also via contaminated bed and body linen.Animal infections or sources of infection are documented in just six cases  . A so-caT. indotineae was initially observed clinically by the fact that the dermatophytosis does not respond to treatment and worsens despite adequate oral antifungal therapy. This may be due to in vitro resistance or at least reduced in vitro sensitivity of the respective dermatophytes [T. indotineae [T. indotineae. In 2017, the first reports of in vitro resistance to terbinafine in India appeared.The terbinafine resistance of tophytes . The tredotineae . It is nT. rubrum isolates were resistant to terbinafine in vitro in a multicenter study in India [T. indotineae no longer respond satisfactorily to topical or oral treatment with this allylamine [T. indotineae is already being seen and must also be expected in the future in countries like Germany and others in Europe and elsewhere [Up to 76% of the isolates of this species and at least 57.1% of the examined in India . It is alylamine . A similThere seems to be an indubitable and oft-considered temporal association of proprietary creams containing topical steroids and antifungal/antibacterial agents with this relatively sudden occurrence of previously uncommon and extensive therapy-refractory chronic widespread dermatophytoses. These creams at times contain as many as four different antimicrobial agents, with three being the norm. These include antimycotic agents , antibiotics  and/or antiseptics  ,44,45. TSteroid combination creams such as those found in India are available in a multitude of countries worldwide, e.g., in the African continent, and in Arab countries. In some countries, e.g., in Germany and other European countries, it is not possible by law to obtain these without prescription, but these countries are the exception.tinea pseudoimbricata [T. concentricum, endemic to the Philippines and Indonesia [T. tonsurans infections, it is seen regularly, if not commonly, in patients of dermatophytosis caused by T. indotineae in India and beyond [T. indotineae often begins as tinea corporis, tinea cruris, or tinea genitalis, sometimes concurrently, as inflammatory or hyperpigmented scaly and severely itchy lesions. Sometimes the lesions are difficult to label as dermatophytosis, but eventually lesions in the groin spread posteriorly to the gluteal region and to the trunk and extremities as direct spread, leading to large lesions , is reported to have superior in vitro activity against zoophilic and anthropophilic dermatophytes [T. indotineae in India [Reduced in vitro sensitivity of strains isolated in Germany to itraconazole has been reported recently . The poitophytes  and espein India .T. rubrum to T. indotineae causing an epidemic-like situation in countries of the Indian subcontinent, especially India, and neighboring countries like UAE, Oman, Iran, amongst others, and now found spreading to Europe, is not only a bothersome disease but has become a public health issue due to the number of individuals affected and the misery it causes. Population-based studies are needed from the Indian subcontinent to understand this disease better. Molecular diagnosis of the incriminated T. indotineae is essential but is not readily available to the vast majority, including large teaching hospitals. The quality of antimycotic drugs, especially itraconazole, needs to be rigorously checked, and finally, strict implementation of laws favoring prescription-only drugs, curbing over-the-counter sales of antifungal drugs, and a strict ban on the manufacture and sale of FDCs containing antifungal agents and potent topical steroids, especially clobetasol propionate, are measures that are the need of the day. Regular discussions with policy makers and bureaucrats involved in ministries of health should be encouraged to act, and plans made with their cooperation should be implemented as soon as possible to curb this menace which is sweeping through several countries and which may reach hitherto unaffected regions by the virtue of migration and tourism, which are bound to increase in the post-COVID era.To conclude, the rather sudden epidemiologic shift from


Dear Sir,Taenia eggs by a woman with the aim of losing weight.Cysticercus bovis, the Taenia saginata metacestode), usually in the form of a pill. The theory is that the tapeworms will reach maturity in the intestine, and that the adult stage will absorb food. This could cause weight loss, along with possible symptoms such as diarrhea and vomiting.We have read with curiosity, as well as with alarm, the article by Zhang et al. on a human case of disseminated cysticercosis because of the intentional ingestion of T. saginata contained in the pills. If the eggs were viable, they would turn into cysticerci, but only in intermediate hosts, cattle in the case of T. saginata. Therefore, these eggs would not have any effect in humans, which was the reason why the authors did not find any reference to the ingestion of Taenia eggs to lose weight. Likewise, the authors speculated that the ingested eggs could be T. solium, rather than T. saginata, as the patient developed disseminated cysticercosis, and among human Taenia tapeworms only the eggs of T. solium are infective for humans.T. asiatica, a species whose ability to cause human cysticercosis is still unknown,T. saginata adult stage) through the morphology of the gravid proglottids, it should be considered that T. asiatica has the same gravid proglottid morphology as T. saginata. Taenia asiatica is present not only in China (where the case is reported) but also in many other Asian countries,T. asiatica cross-reacts with T. solium in the ELISA the authors used.T. asiatica was responsible should not be ruled out.According to the case report, the patient did not ingest any cysticerci, but rather the eggs of T. solium eggs for human consumption, considering the risk of dying of neurocysticercosis, as happened in an illustrative case with death after the intentional ingestion of T. solium eggs published in 1994.T. solium, to prevent possible fatal cases of neurocysticercosis in other pill consumers.It would be hard to believe that the providers of the pills purposely supplied


Mycobacterium (M.) avium complex; M. fortuitum; and other NTM species, including M. marinum infection, known as fish tank granuloma, and M. ulcerans infection, which is described as a Buruli ulcer. NTM could be considered as water-borne, air-borne, and soil-borne pathogens (sapronoses). A lot of clinically relevant NTM species could be considered due to the enormity of published data on permanent, periodic, transient, and incidental sapronoses. Interest is currently increasing in mycobacterioses diagnosed in humans and husbandry animals (esp. pigs) caused by NTM species present in peat bogs, potting soil, garden peat, bat and bird guano, and other matrices used as garden fertilizers. NTM are present in dust particles and in water aerosols, which represent certain factors during aerogenous infection in immunosuppressed host organisms during hospitalization, speleotherapy, and leisure activities. For this Special Issue, a collection of articles providing a current view of the research on NTM—including the clinical relevance, therapy, prevention of mycobacterioses, epidemiology, and ecology—are addressed.Nontuberculous mycobacteria (NTM) represent an important group of environmentally saprophytic and potentially pathogenic bacteria that can cause serious mycobacterioses in humans and animals. The sources of infections often remain undetected except for soil- or water-borne, water-washed, water-based, or water-related infections caused by groups of the  The first comprehensive piece of information on the ecology of mycobacteria came from a book by Jindrich Kazda that was published in 2000 . The secIt can be assumed that due to changes in the environment, and especially the permanent decline in the healthy resistance of the human population , mycobacteriosis caused by nontuberculous mycobacteria (NTM) will present an ongoing issue. In addition, improvements in water treatment, namely, the introduction of widespread water disinfection (chlorination), have provided a selective environment for NTM. New findings on the ecology of NTM have revealed previously neglected or unknown facts related to their long-term survival in various components of the environment, their ability to colonize new substrates, and their interaction with humans. Finding new, abundant, or yet undiscovered sources of some clinically important NTM species will help to more effectively prevent exposure to predisposed individuals. Preventing the exposure of NTM to susceptible individuals is currently one of the most important ways to prevent mycobacteriosis.The persistent incidence of pulmonary and extrapulmonary mycobacteriosis in children and adults is described herein for 2020–2022. Pulmonary mycobacteriosis is severe in individuals with a predisposition to infection and in individuals with comorbidities in the United States ,24, and Mycobacterium (M.) mageritense infection [M. marinum hand infection masquerading as tinea manuum [M. marinum infections in 40 patients in the Netherlands [M. terrae and M. intracellulare tenosynovitis following hurricane relief efforts in the USA [M. smegmatis skin infection following cosmetic procedures [M. abscessus outbreaks at pediatric dental clinics were documented with more than 20 confirmed infected children with a median age of 6 years in both clinics in Atlanta, Georgia [M. szulgai infection was diagnosed in a 25-year-old female with systemic lupus erythematosus graduating in veterinary medicine in Portugal [Extrapulmonary mycobacterioses have been described primarily as skin infections. The following species of NTM have been demonstrated: tattoo-associated cutaneous nfection , M. maria manuum  and the herlands , M. terr the USA , and M. ocedures . Two M.  Georgia . DissemiPortugal .There are reasons for diligently seeking NTM resources for patients. The review article from Faverio et al. shows that the common aim in Italy is a comprehensive approach to the treatment and prevention of pulmonary mycobacteriosis. In addition to antibiotic treatment, it seeks the following four preventative steps: (1) avoiding exposure to environments colonized with NTM and evaluating the lifestyle and habits of patients; (2) implementing a personalized pulmonary rehabilitation plan and airway clearance techniques to improve symptoms, exercise capacity, health-related quality of life, and functional capacity in daily living activities; (3) evaluating the patient’s nutritional status, intervening to improve their health-related quality of life and control gastrointestinal side-effects during antimicrobial therapy ; and (4) managing comorbidities affecting disease outcomes, including structural lung diseases, immune status evaluation, and psychological support . As the M. tuberculosis is primarily transmitted by person-to-person contact. In contrast, NTM are mainly transmitted from the environment [p < 0.001). The following parameters were significant in terms of the personal characteristics of patients: female sex (p < 0.001), age (p < 0.001), and HIV/AIDS incidence (p < 0.001) [ironment ,2. Due t< 0.001) .M. avium complex isolates [M. avium complex isolates from human patients with lung and disseminated infection from animals and the environment. In particular, isolates from humans and animals differed. Similarly, this has also been demonstrated in isolates from human patients and their environment [Sophisticated molecular biology methods have been used for the clinical and environmental epidemiology of isolates . In the ironment . In the ironment .In 2020–2021, a survey of the Czech market was conducted on plant fertilization products that contained peat and/or bat guano. We found a high incidence of NTM that contaminated up to 80–100% of these products . FurtherIn many known and newly described mycobacterial species, we often encounter only a vague statement that their source for humans or animals is “most likely” the environment. In the “better cases”, at least the component of this environment  in which the species “most likely” occurs is mentioned. It is not at all easy to find comprehensive knowledge about the ecology of the exact mycobacterial species that often causes serious human and animal diseases. There are few scientific teams that have systematically studied the ecology of mycobacteria. These teams usually carry out research with the intensive cooperation of physicians and veterinarians, biologists, zoologists, ecologists, climatologists, pedologists, microbiologists, biostatistics, molecular biologists, and other experts. In this way, they succeed in connecting these relatively “closed parallel highly specialized worlds”.Humans have created ideal habitats for the NTM, particularly via plumbing. Specifically, the following characteristics of plumbing have been selected to increase the presence of NTM: surfaces that promote adherence and biofilm formation; disinfection to kill off microbial competitors; heating, which increases the numbers of NTM; and stagnation, under which NTM can grow while other pathogens may not.Therefore, this Special Issue presents a number of results, opinions, and conclusions regarding various environmental matrices in relation to mycobacteria.M. fortuitum was first described as a new species in 1938. Since then, a total of 16 members of the M. fortuitum group has been described. Their occurrence in aquarium freshwater and marine fish can be found in the article by Mugetti et al. [M. fortuitum group.i et al. . Their ci et al. ,45. All M. chimaera in the aquatic environment, especially in water pipes and cooling tanks of cardiology units, currently represents a significant health risk. Its capture is described in the publication by Zoccola et al., which concerns the validation of a novel diagnostic approach combining the VersaTREK™ system for recovery and real-time PCR for the identification of M. chimaera in water samples [The occurrence of  samples .M. algericum, M. bohemicum, M. duvalii, M. europaeum, M. fallax, M. goodii, M. hassiacum, M. hiberniae, M. malmoense, M. mucogenicum, M. nebraskense, M. numidimassilliense, M. parmense, M. porcinum, M. saskatchewanense, M. simiae, and M. timonense. Especially surprising was the high concentration of M. avium ssp. hominissuis, M. malmoense, and M. xenopi in peat and plant material. The detection of mycobacterial DNA with qPCR from live and dead mycobacterial cells greatly accelerated and facilitated the detection of the NTM load in various environmental matrices.The cultivation evidence of NTM from heavily contaminated environmental matrices was often accompanied by the contamination of culture media. This greatly hindered the capture of different NTM species, which were present in different matrices. The publication by Ulmann et al.  presentsM. fortuitum group, M. chelonae group, M. mucogenicum group, and M. avium complex) and NTM species belonging to risk group one  were isolated from guano for the first time [Based on the results published in this Special Issue, bat guano can be considered a significant source of NTM. High culture positivity and the considerably wide spectrum of NTM species in guano originating both from caves (cave guano) and from summer colonies from attics (attic guano) is necessary in European conditions , considered as highly interesting. Many species from risk group two  and NTM belonging to risk group two  is interesting. Indeed, dust is not very often considered to be a significant source of NTM compared to water and water biofilms [From another publication by Hubelova et al. , it is pbiofilms ,2.Two reviews published in this Special Issue open novel perspectives on the ecology of NTM ,51. The The second review, written by Pavlik et al. , comes uThe purpose of these articles is not to tire or bore the reader with data. These researchers show new methods of detecting mycobacteria, new perspectives on the importance of the environment in their survival and spread, and the current understanding of the ecology of mycobacteria. We, therefore, hope that these published data will also serve as a guide for further thinking about the ecology of mycobacteria.


Plodia interpunctella (Hübner) is an important grain storage pest in China that is distributed in almost every province and that has caused huge economic losses. In this study, we adjusted the biological parameters of the Indian grain borer and then used the CLIMEX model to predict the detailed potential distribution of P. interpunctella in China under current and future conditions. Under historical climatic conditions, the central, northeastern, and southern regions of China are highly suitable habitats for it. Because of temperature change in the future climate, suitable habitats will increase in the eastern part of Qinghai and will decrease in the mid-eastern, northeastern, and southeastern parts of China. This study provides important information for controlling the further spread of the Indian corn borer in China.Plodia interpunctella (Hübner)  is a notorious stored-grain pest that can be found in most parts of China. The corpses, excretions, and other secretions of P. interpunctella larvae cause serious grain pollution, seriously affecting the nutritional and economic value of stored grain in China. To elucidate the potential distribution of P. interpunctella in China, we used the CLIMEX 4.0 model to project the potential distribution of the pest using historical climate data (1960–1990) and estimated future climate data . Under the historical climate situation, P. interpunctella was distributed in most areas of China, and its highly favorable habitats account for 48.14% of its total potential distribution. Because of temperature change in the future climate, suitable habitats will increase in the eastern part of Qinghai and will decrease in the mid-eastern, northeastern, and southeastern parts of China. Under these scenarios, the area of this pest’s highly favorable habitat will be reduced by 1.24 million km2, and its proportion will decrease to about 28.48%. These predicted outcomes will help to distinguish the impact of climate change on the potential distribution of P. interpunctella, thereby providing important information to design early forecasting and strategies to prevent pest harm to stored grain.The Indian meal moth  Plodia interpunctella (Hübner) is a primary injurious insect to raw and processed stored food products [P. interpunctella can migrate and spread via infected food goods, which is accelerated due to the development of globalization, leading to it spreading wherever there is permanent human habitation [P. interpunctella harms more than 170 species in storage, including various grains, dried fruits, pulses, dried vegetables, etc. [P. interpunctella has caused great economic losses in the United States, China, Korea, Ethiopia, and in other countries [The Indian meal moth products ,2. The fproducts ,4,5. Becbitation ,7. P. ines, etc. . After tountries ,10,11,12P. interpunctella is widely distributed and occurs with high frequency in China, and with the warming climate, it has multiplied in the high-latitude regions of China in Alar, Xinjiang [P. interpunctella in China mainly focuses on its molecular biology [P. interpunctella in China. Understanding its potential future distribution pattern will enable the governmental pest management agencies to implement corresponding measures to prevent it from continuing to cause losses.Xinjiang ,14. The Xinjiang . RelevanXinjiang . ClimateXinjiang . The res biology ,19, sex  biology , and con biology ,22, whilThe potential geographical distribution of different invasive species has been shown to be primarily evaluated using ecological niche models, such as ANUCLIM/BIOCLIM, CLIMATE, CLIMEX, DOMAIN, GARP, HABITAT, and MaxEnt . Among tP. interpunctella in China under future climate conditions. We adjusted parameter values according to the published literature to better fit the known distribution in China [P. interpunctella in China under historical and future climatic conditions is helpful for relevant pest management departments to implement early warning and prevention policies.There is little research on the detailed potential distribution of in China . In thisThe CLIMEX model is a dynamic and bioclimatic niche simulation model that uses species parameters and climate data to predict the potential geographical distribution of species . In thishttps://www.climond.org/ (accessed on 4 November 2021)) gridded 10’ spatial resolution meteorological data were used for modeling the distribution of P. interpunctella in CLIMEX because of its good spatial resolution [P. interpunctella in the future climate change scenarios predicted for 2030, 2050, and 2070 [CliMond , the Global Biodiversity Information Facility (GBIF), and the published literature reviews related to biological characteristics, as well as scientific information on Web of Science, Science Direct, China National Knowledge Internet (CNKI http://cnki.net/ (accessed on 23 December 2021)), PubMed, and ASP. Up until now, the P. interpunctella has been distributed in most parts of China, except in areas with a harsh climate.Records detailing the distribution of P. interpunctella feeds on different diets, differences can be observed in its biological parameters. Rice is the staple food of most people in China and is stored in every household [P. interpunctella to rice. Therefore, we chose to set the relevant biological parameters of P. interpunctella as the standard when it fed rice. Jung [P. interpunctella is higher when the temperature range is 20–28 °C, with growth inhibition being observed at 32–35 °C [P. interpunctella died within 15 d of being at 30 °C [When ousehold . China rousehold , so it wce. Jung  found th32–35 °C . Lee’s eat 30 °C . Based oP. interpunctella. When fresh vegetables and fruits are dried, they emit fragrances and are easily infected by P. interpunctella. Therefore, combined with the lowest default value provided by the CLIMEX template, the lower soil moisture threshold (SM0) was set to 0.05 to indicate that P. interpunctella can continue to develop in dry environments. The lower optimal soil moisture (SM1) was set to 0.2, representing relatively dry soil, to match the distribution records of P. interpunctella in northwest Xinjiang, China. The widespread distribution of P. interpunctella in Shanxi, Henan, and other northern provinces shows that it can grow well in temperate climates. Therefore, according to the template in the software and Jung’s research [There are few moisture index studies related to research , we alsoP. interpunctella is unable to survive at temperatures above 35 °C.Heat Stress: The heat stress threshold temperature (TTHS) was estimated to be the same as DV3 (35 °C), and the heat stress temperature rate (THHS) was estimated to be 0.0048; P. interpunctella larvae by Feng et al. [P. interpunctella in the southwest part of Liaoning province and in the midwest part of Inner Mongolia [Cold Stress: Using information from a supercooling point study of g et al. , the colMongolia .P. interpunctella, the relevant parameter values could not be estimated. Recent surveys show that the frequency of the P. interpunctella in northern China is significantly higher than that in southern China, with the highest frequency in Shanxi Province [Dry Stress and Wet Stress: Due to the lack of available information on the dry and wet stress of Province . Shanxi Province . We thusMoisture parameters without units are dimensionless indices of soil moisture .P. interpunctella in different areas of China more intuitively, we refer to its actual distribution in China and Lee’s research [In order to present the suitability of research  and diviP. interpunctella in China. From the distribution map generated by ArcMap, we can calculate the total area of the potential distribution of P. interpunctella in China to be 7.00 million km2; this represents 72.92% of the total area of mainland China. The area comprising a highly favorable habitat was 3.37 million km2, which is 35.11% of the total area of mainland China and accounts for 48.14% of the total area of potential distribution. This potential distribution includes most of the area of Heilongjiang, Jilin, Hebei, Ningxia, Guangdong, Guanxi and Yunnan, Liaoning, Beijing, Tianjin, Shanxi, Shaanxi, Shandong, Henan, Guangdong, Guanxi, Yunnan, the southeast of Gansu, the east Sichuan, the north of Jiangsu and Anhui, the south of Guizhou and Fujian, the northeast of Inner Mongolia, the southwest Taiwan, and a small area of Tibet.The model prediction results show that, except for Alar, the recorded distributions collected from CNKI were within the potential distribution area predicted by CLIMEX . This pr2, which is 18.54% of the total area of mainland China and accounts for 25.43% of all the area of potential distribution. The distribution of this habitat covers southern Hubei, Jiangsu, and Anhui; most areas of Hunan and Jiangxi; northern Guizhou, Inner Mongolia, Fujian, and Taiwan; and small areas of Hebei, Heilongjiang, Jilin, Qinghai, Gansu, Yunnan, Guangdong, Guangxi, and Tibet. The area of the marginally favorable habitat was 1.85 million km2, which is 19.27% of the total area of mainland China and accounts for 26.43% of the entire region of potential distribution. The main areas belonging to this habitat were the northwest of Gansu, Xingjiang, and Qinghai; the middle part of Hunan, Inner Mongolia, and Zhejiang; northern Yunnan; western Sichuan; and southeastern Tibet.The area of the favorable habitat was 1.78 million kmP. interpunctella in China under future climate conditions is shown in The potential distribution range of P. interpunctella in the six major agricultural provinces of China [P. interpunctella for a long time. By 2070, except for Jilin, the unfavorable habitat areas in five other provinces had begun to increase. The highly favorable habitat areas of Anhui, Henan, and Shandong decrease year by year, and those of Inner Mongolia, Heilongjiang, and Jinlin increase and then decline. P. interpunctella in China over time. The total region of potential distribution and the area covered by marginally favorable habitat will increase, while the district covered by highly favorable habitat will show a marked decline, with the favorable habitat changing very little. The area of the total potential distribution is observed to increase by 480,000 km2 from 1990 to 2070. The increased region of marginally favorable habitat is the largest, covering an area of 1.86 million km2. The reduced area of the highly favorable habitat was 1.24 million km2.of China . Before The subtraction of the difference data between three sets of future climate EI values and historical climate EI values was interpolated into ArcMap, generating EI interpolation variation maps . We founP. interpunctella survival in China during different periods from 2030 to 2070, the climatic favorability levels increase in the southeast of Liaoning, Tibet, and Jilin; the northern regions of Yunnan; the southern regions of Sichuan and Gansu; and most areas of Guizhou and Taiwan, and the EI value increased from 1 to 40. The EI values for the north of Sichuan; the eastern part of Qinghai; and the junction of Gansu, Qinghai, and Sichuan would increase the most, by up to 40. Climatic favorability levels for P. interpunctella would decrease in the northern, southern, and east-central parts of China, with decreasing EI values ranging from 1–32. EI values for the southwest of Guangdong, most of Guangxi, and the northern part of Hainan would decrease the most, reaching 42.By comparing the changes favoring P. interpunctella has the characteristics of euryphagy and crypticity, and it is distributed all over the world, which indicates that it has a strong continuous transmission ability. Therefore, we predicted its potential distribution under the current and future A2 SRES climate scenarios in China. Among the cities in China where the distribution of P. interpunctella is known, except for Alar, Xinjiang, the rest are within our forecast range, which proves that our model is reliable.The development of models for predicting the potential distribution of insects in different climatic conditions makes the prediction results more accurate and can provide valuable data for the monitoring and control of harmful insects ,46. CLIMP. interpunctella negatively in the central-eastern and southern provinces of China. In general, the total area suitable for this pest will decrease by 2070. It should also be noted that northwestern China will become more suitable. The global climate model (GCM) CSIRO-Mk3.0 (CS) data under the A2 SRES scenario used in this study predict that the global climate is warming at a rate of 0.5 °C per decade and that precipitation will decrease [P. interpunctella cannot survive above 35 °C and that high temperatures inhibit its growth, with a survival rate of only 1% when the temperature was 35 °C [P. interpunctella.The predicted climate changes will affect decrease . Previouas 35 °C ,31,48. Ias 35 °C . By 2070P. interpunctella around the world according to four different diets. Jung indicated that rice production areas, such as Hunan and Jiangxi, are high-risk areas that are susceptible to infestation by the P. interpunctella, and our findings support this conclusion. Jung predicted that only Henan province in northern China has EI values that indicate that it could be infested by the P. interpunctella, while our model showed that most of the provinces in northern China had high EI values. The report shows that, among the different grain-producing areas of the 11 provinces in China’s main grain storage ecological areas, the occurrence frequency of P. interpunctella is the highest in Shanxi, Liaoning, and Henan in the north, with occurrence frequencies of 42.3%, 39.5%, and 36.8%, respectively [P. interpunctella in China. Because the climate data are from the same source, the differences in the biological parameter settings lead to differences between the two research results. In recent years, the discovery sites of P. interpunctella in China have continued to increase. Based on its biological parameters, such as its supercooling point and current distribution, the parameters have been adjusted. Finally, the EI values calculated by the model may be different, which makes the specific prediction distribution range different.Before this study, Jung et al.  predicteectively , which pP. interpunctella to survive under such conditions. The Tarim Basin in the south of Xinjiang has a higher distribution of desert, low vegetation coverage, perennial natural disasters, such as sandstorms and dry–hot wind, and a high dry stress index, which is also not suitable for the growth of P. interpunctella. In recent years, the Zhungeer basin in the north of Xinjiang and the Qaidam basin in the northwest of Qinghai have experienced changes in their local hydrothermal conditions due to the continuous transformation of the local agricultural ecosystem [P. interpunctella. The plain in the northern and hilly areas in the south is mild and suitable for planting, and most areas are in a highly suitable living environment for P. interpunctella, making it extremely vulnerable to infection.China’s terrain has gradually increased from the eastern coast to the western inland area and is arranged in stepped shape . The avecosystem . Both haP. interpunctella harms stored grain, its biological characteristics will change with time due to the different positions and environments of granaries, and these factors may also affect the predicted distribution. The adults of P. interpunctella have strong mobility, which can infect storage within the range of 21~276 m, and its dispersion ability also has an important influence on the distribution [P. interpunctella will be further explored. In a subsequent study, we will also continue to investigate the distribution of the P. interpunctella in China, explore the specific effects of external environmental changes in the biological parameters of the P. interpunctella, and establish a model evaluation system to improve model performance.CLIMEX model predictions are primarily based on climatic factors, but nonclimatic factors, such as biological interactions, genetic evolution, graphic barriers, and human activities, might affect the studies of species distribution . By moniribution . Recent ribution . With thP. interpunctella. Since the beginning of the 21st century, grain production in northern China has increased substantially and has been shipped to the south in large quantities, directly promoting the construction of grain logistics node layout, logistics parks, and grain storage warehouses [P. interpunctella are small, so it is easy for it to spread with the host for a long period time. If no active control measures are taken, the spread of P. interpunctella in China will continue to accelerate. The intelligent detection and early warning technology of P. interpunctella can determine its dynamic changes in a timely manner and accurately provide a powerful guarantee for preventing it from infecting stored grains. Near-infrared spectroscopy (NIR) coupled with discriminant and modeling classification methods can be used to detect the presence of P. interpunctella and is not destructive to the tested grains [P. interpunctella, it can be used to control gamma radiation, gaseous ozone, and high nitrogen [P. interpunctella in large commodities. However, the extensive use of methyl bromide has been proven to cause damage to the ozone layer, and it is being phased out [The model in this study shows that in the next 50 years, most of China will be a potentially suitable distribution area for rehouses . The ward grains . Metal sd grains . Once grnitrogen ,65,66. Onitrogen . The cheased out ,69. Phosased out . Studiesased out . Extractased out .P. interpunctella in China under historical and future climate data using the CLIMEX model. The results show that in the next 50 years, most areas of China are potential distribution areas for P. interpunctella, except for parts of the Qinghai–Tibet Plateau and Tarim Basin. With the warming of the climate, the expanding areas are mainly located in the eastern part of Qinghai, while the mid-eastern, northeastern, and southeastern parts of China appear likely to become unfavorable areas. China should continue to implement strict monitoring, prevention, and control measures for P. interpunctella to prevent its spread all over the country and to achieve the goal of ensuring food security.In this study, we analyzed the potential distribution of


Blattisocius mali is a promising predatory mite for pest control, and a thorough understanding of its dispersal pathways in crops could be crucial for successful plant protection. So far, it has been reported as transported by several species of drosophilids. Our research indicates an ectoparasitic relationship between this species and drosophilid fruit flies. We used the flightless D. melanogaster and D. hydei commercially raised as live pet food. Female mites not only attached to flies but also fed on them, and their presence resulted in an increase in fly mortality. Although both fly species used similar defense tactics, mites had more difficulties getting onto D. hydei. Whether the wild D. hydei and D. melanogaster can also transport B. mali will be shown by further laboratory and field studies.There are a number of reports on the dispersal of predatory mites on insects, although much less is known about their mutual interactions and possible relationships. While some mite species use insects only as means of transport, others also feed on them, significantly lowering the carriers’ fitness. Blattisocius mali is a promising biological control agent which has been reported as transported by several drosophilid species. Our goal was to determine the type of relationship between this mite and fruit flies. We used flightless females of Drosophila melanogaster and D. hydei, which were commercially raised as live pet food. The predatory females mostly attacked the tarsi of the flies and then preferentially moved to the cervix or close to coxa III, where they eventually drilled their chelicerae and started feeding. Although both fly species used similar defensive tactics, more B. mali females did not attack D. hydei or did so with a delay, and a higher percentage of mites fell off the D. hydei tarsi during the first hour of observation. After 24 h, we noted the increased mortality of flies exposed to the presence of mites. Our study indicates the ectoparasitic relationship of B. mali with drosophilids. However, further research is needed to confirm the transport of this mite on wild D. hydei and D. melanogaster, both in the laboratory and under natural conditions.Predatory mites dispersing by means of insects are often ectoparasites and may use various tactics to get onto the host, counteract its defenses, and diminish its survival.  Poecilochirus carabi G. Canestrini and R. Canestrini on the burying beetle Nicrophorus vespilloides Herbst [Parasitellus fucorum (De Geer) on bumblebees Bombus sp. [Macrocheles muscaedomesticae (Scopoli) and M. subbadius (Berlese) [Spreading using insects is a common mode of dispersal in mites ,2,3,4,5.s Herbst  or Parasmbus sp. , others Berlese) ,11. MoreBerlese) . DespiteBerlese) , there iBlattisocius mali (Oudemans). It is a promising biological control agent for acarid mites, eggs of the potato tuber moths, and nematodes [Over a dozen mite species are reported as associated with drosophilid flies, both in the wild and in laboratory cultures ,16,17,18ematodes ,23,24,25Blattisocius mali belongs to Blattisociidae (Mesostigmata), a family of cosmopolitan predatory mites found both in the soil and in litter, on plants and stored products, and is frequently associated with rodents, insects, and birds [Blattisocius Kegaan, B. keegani Fox, B. dendriticus (Berlese), B. patagorium Treat, and B. tarsalis (Berlese) have been reported as transported by noctuiid and pyralid moths [B. patogorium and B. tarsalis, an ectoparasitic relationship with moths has been suggested [B. apis Basha and Yousef and B. trigonae Radhakrishnan and Ramaraju, were found on the body of honey bees, while B. capsicum Basha and Yousef was found in association with juveniles of psocopterous species inhabiting stored hot peppers [nd birds . Among sid moths ,27,28,29uggested . Others, peppers ,32.B. mali has been recorded only in fruit flies (Drosophilidae) from both the Drosophilinae and Steganinae subfamilies. In Finland, the mites  have been found on Drosophila littoralis Meigen, D. montana Stone, D. ezoana Takada and Okada, and D. lummei Hackman [B. mali has been attached to D. hexastigma Patterson and Mainland [Phortica semivirgo Máca. Blattisocius mali has also been reported as associated with drosophilids in laboratory cultures [So far,  Hackman . In the Mainland . FinallyMainland  found fecultures .B. mali seemed to be inserted through the integument of fruit flies, which suggests that the association of this mite with drosophilids could be parasitic.Phoretic mites attach themselves to insects in a variety of ways ,5,33,34.B. mali and drosophilid flies. First, we examined the behavioral tactics the mite uses to get onto the body of the flies, where it preferentially attaches itself, and what the influence of the drosophilid species could be on this process. Secondly, we checked whether B. mali feeds on the fruit flies during transportation, and thirdly, whether the dispersal of the mite with the aid of the flies affects the survival of its carriers.The aim of this study was to determine the type of relationship between B. mali and the mold mite Tyrophagus putrecentiae (Schrank) came from the laboratory mite cultures maintained in the Section of Applied Entomology, Department of Plant Protection at Warsaw University of Life Sciences (WULS). Blattisocius mali was identified by Dr. K. Michalska using the key by Karg [Blattisocius mali was mass-reared on various stages of T. putrescentiae in wheat bran on soaked foam platforms (22 cm × 15 cm × 2.5 cm), which were covered with foil and placed within broader vessels (50 cm × 70 cm × 20 cm) filled up with water. The cultures of B. mali were maintained in a climatic room, at 21–23 °C, with a photoperiod of 16/8 h (L/D). The population of T. putrescentiae was fed with instant yeasts and maintained in a desiccator  at 26 °C and 80–85% RH in darkness.The stock population of  by Karg  and subsD. melanogaster Meigen and D. hydei Sturtevant  and females of D. melanogaster (n = 30) or D. hydei (n = 30). Apart from the treatment combination, a control was prepared in which female flies of D. melanogaster (n = 30) and D. hydei (n = 30) were kept singly, without mites. Observations were made at 5–50× magnification using a stereo microscope  connected to a cooled light source. The observational chambers were constructed from colorless, transparent 200-μL Eppendorf pipette tips  that were 3 cm long. The chamber had a vent (ca. 0.5 mm in diam) at the narrower end, and at the broader end, it was plugged with a piece of cotton wrapped in gauze. First, single fruit fly females were introduced into the chamber, and then , the single 24-hour-starved females of B. mali were released into it. In contrast to fully satiated mites, the idiosoma of hungry mites was flat and brown. The mites were transferred with the aid of a fine brush, while the fruit flies were transferred using an aspirator (handmade). The mites were starved in isolation within a conical chamber of glass cages (4 cm × 3 cm × 0.3 cm) following the methods described by Robertson [The tests were carried out within small observational chambers using pairs of randomly selected females of obertson . The conB. mali attachments to D. melanogaster and D. hydei were conducted. We regarded the place of location of the mite’s mouthparts on the fly’s body as a site of the mite’s attachment. For the description of the flies’ thoracic morphology, we used the nomenclature from Fabian et al. [B. mali females fed on fruit flies and whether the flies defended themselves against them. The mite’s feeding on the fly was judged by the degree of filling of its idiosoma. We arbitrarily assumed three degrees of the mite’s idiosoma filling: flat, partially filled, and full. The times of particular mite behaviors were measured using a stopwatch.The experiment was divided into two parts. In the first part, which lasted one hour, detailed ethological observations and time measurements of the n et al. . The paiB. mali may remain attached to a fly and feed over a longer period of time and how this may affect the viability of the fly and the mite itself, observations were continued for the next 23 h. After the completion of 1-h observations, the position of the mite on the fly’s body, the degree of the filling of its idiosoma, and the viability of the flies and the mite were noted, and then the observational chambers with the flies and B. mali females, as well as the control flies, were placed in an incubator with a temperature of 23° C and a 16/8 h (L/D) photoperiod. Additionally, we kept the relative humidity in the incubator at 60–70% RH, which, as investigations by Shaffer et al. [B. mali females that stayed with flies, which were either living or already dead, and could be either attached to or separated from the flies’ bodies.In order to examine whether r et al.  and Kamar et al.  have shoB. mali individuals, the analysis took into account the structure of variance and covariance of AR1. An overall assessment of differences in observations expressed as percentages was made using the chi-square test for contingency tables, while pairwise comparisons between observations were made using the chi-square test of independence. Data were presented as mean ± standard error (SE).The statistical analyses were performed using R 4.2.1 software . In the D. melanogaster females showed markedly greater mobility than D. hydei in the first 10 min, even before the mite was placed into the chamber. These females very quickly moved from one end of the chamber to the other, seeking an exit. In the following minutes, D. melanogaster flies, both from the control and the treatment combination, were more often seen in the narrower end of the chamber, where there was a vent. When squeezing into the chamber tip, they curled their legs under them. Drosophila hydei females behaved differently. After one or two attempts to squeeze into the chamber tip, they moved towards the wider end, and when stopped, they remained in a standing position with straight legs.The behavior of D. melanogaster and D. hydei females responded to the mite attack and could make several different attempts to get rid of the mite. When B. mali attached the chelicerae to the tarsus of the fly, then the fly often curled her leg up under her or began to run. To get rid of the mite, she flicked with her tarsus and hit it against the walls of the chamber. Alternatively, she began by grooming and attempted to remove the mite using a leg of the same or one of the other pairs. Similarly, when B. mali stuck its chelicerae in the wing, thorax, or abdomen, then the flies tried to throw the mite off, rubbing the places where the mite had attached intensively with their legs I, II, or III. The attacked fly could also jump or roll over on her back, waving all her legs intensely.Both D. hydei than in D. melanogaster . In D. melanogaster, only one trial out of 37 attempts to remove B. mali was successful (2.7%), while in D. hydei, 8 out of 21 trials (39%) were completed with the removal of the mite. By contrast, the percentage of successful attempts to throw B. mali off other body parts did not differ significantly between fly species . Drosophila hydei females managed to remove the mite 4 times out of 8 attempts (50%), while D. melanogaster only once in 6 attempts (16.67%).During the first hour of observation, the percentage of successful attempts to throw the mite off the tarsus was significantly higher in B. mali getting onto D. melanogaster or D. hydei and its final attachment to their body. The first one was attack. During this phase, the mite most frequently attached to the tarsus of the fly leg, rapidly sticking its chelicerae into the first, second, or third segment of the tarsus. Only when the fly ‘calmed down’ (stopped and did not wave its legs) did the mite begin to climb up the fly’s body. Either (1) it moved along the more or less straight leg, onto the cervix (neck), thorax, or abdomen, or (2) if the fly had curled its leg under her, it moved from the tarsus or another part of the leg which was touching or a short distance from the fly’s body, directly onto the fly’s body. In the latter case, the predator stretched legs I or legs I and II in front of him and hooking them on the protruding part of the fly’s body, passed onto its cervix, thorax, or abdomen. (3) B. mali also got on drosophilids when body parts other than the tarsus were in contact with the wall of the observation chamber or the chamber plug at its wider end. In this case, the mites stuck their chelicerae into the thorax, abdomen, or wings and much more rarely into the head or mouth parts of the flies  was noted  or nearby, and when the fly stopped there, the attack began. The predators could attack the fly from the front  or from the back , or from any other side if it happened to be close to the contact point of other parts of the fly’s body with the wall of the chamber.as noted . Not infThe passage of the mite from the site of attack to the site of the final attachment on the fly’s cervix, thorax, or abdomen was mostly very slow, as if the mite were ‘creeping’, and took place with or without stops. The final attachment was preceded by a characteristic drilling of the mouthparts into the integument of the insect. After inserting the chelicerae, predators raised the idiosoma upwards and began waving legs IV, III, and even II and turning the body sideways intensively. It was only after some time that the predators stopped waving their legs, their bodies settled on the flies ‘for good’, and they became motionless.D. melanogaster females, while in the case of D. hydei, 23.33% of B. mali females showed no interest in the insects . The mite attacked the D. melanogaster fly after, on average, 479.77 ± 80.31 s from the moment of release to the chamber, while attacking D. hydei after a significantly longer time, on average after 729.09 ± 113.21 s . We observed, in total, 31 mite attacks (and first attempts to attach) on D. melanogaster and 32 mite attacks on D. hydei. However, in the case of D. hydei, the percentage of failed attempts to attach at the attack site was significantly higher, at 53.13%, while in the case of D. melanogaster, it was only 6.45% . The mite failed to attach to the tarsus, rim of the wing, head, or tip of the abdomen of D. hydei, and it was usually thrown off these sites by a defending fly (13 attempts), much more rarely dropping off by itself (4 attempts). During the attacks on D. melanogaster, the mite attempted to attach to the tip of the abdomen. Once, it was thrown off by a fly, and once, it fell off by itself.During the first hour of observation, all the tested female predators attempted to attack 2 = 11.73, df = 1, p = 0.0006). A total of 25 out of the 30 B. mali females kept with D. melanogaster (83.33%), and only 11 out of the 30 females (36.67%) kept with D. hydei got onto the cervix, thorax, or abdomen of the fly, where they finally drilled into its body. There were significant differences in the percentage of mites entering the body of D. melanogaster, either from the tarsus of a straight or curled leg or from the junction of the other fly’s body parts with the chamber wall . Most B. mali females entered the body of D. melanogaster by moving from the tarsus, especially from that of the curled leg . However, in comparison with D. melanogaster, a much greater percentage of mites entered the D. hydei body by using the contact of body parts other than the fly’s tarsus with the chamber wall   .B. mali to finally attach to the fruit flies and also the time spent in each location phase varied greatly among mites , and to D. hydei was 1193.46 ± 326.07 s  for D. hydei, and did not differ significantly between mites attacking either fly species . The fly species also had no effect on the average time the predator spent in a given phase of location on the fly’s body , and no significant interaction between the fly species and the location phase was detected  . The mean time spent by a predator in the phase of attack and first attachment was significantly longer than that spent during the passage from the attack site to the site of final attachment (p ≤ 0.05) or during the drilling phase (p ≤ 0.05), both in D. melanogaster or D. hydei (p > 0.05).The total time it took for ng mites . The ave 0.2507) . HoweverD. hydei . In neitB. mali females moving from the attack site could have attached to flies in various places all over their body, but eventually, they began to drill with their chelicerae only in one of them. In D. melanogaster, there were 4 such sites on the cervix, thorax, and abdomen, while in D. hydei, there were 8 sites  and 24 h . In D. hydei, however, the percentage of mites drilled into the cervix or near the coxa III during the first hour was similar to other places on the thorax or abdomen . Only for the total records of attachment, obtained during 24 h, were there significant differences in the frequency of B. mali females attached to the various sites on D. hydei , with a distinct predominance of females stuck, similarly as in the case of D. melanogaster, to the cervix and near the coxa III  or coxa III of D. melanogaster  differed significantly depending on whether the mite attacked the tarsus I, II, or III or other sites of the fly’s body in contact with the observational chamber. The greatest percentage of mites reached the cervix of D. melanogaster by climbing from the tarsus I, while the greatest percentage of mites reached the fly’s coxa III by climbing from her tarsus III. A similar analysis was not performed for D. hydei due to insufficient sample size  or D. hydei   a,b.D. melanogaster and three cases on D. hydei. Only when B. mali females attached to the fly cervix, thorax, or abdomen did their idiosomas gradually fill. Initially, only a thickening of the back of the body, in the area of histerosoma, could be observed in the feeding mites. In the case of mites attached to D. melanogaster, a partial filling of the idiosoma was recorded in the second hour of observation. At this time, the idiosoma of one mite was already completely full. On D. hydei, the first cases of B. mali with completely filled idiosomas were not recorded until the third hour. The mites with full idiosomas were bulky, light brown on the dorsal side, and yellowish on the sides. Interestingly, after 24 h, most of the females feeding on D. hydei had completely filled idiosomas, while on D. melanogaster, partially filled idiosomas.In the 24-h-starved mites, the body was flat and dark brown. They remained flat also when they became stuck to the tarsus of the leg or to the wing and failed to get onto a fly body. Two such cases have been observed on 2 = 3.987, df = 1, p = 0.0354). The mortality was much higher in D. melanogaster than in D. hydei, both with B. mali  and without the mite , and no significant influence of the mite on survival of D. melanogaster was detected   .D. hydei, after 24 h, in combination with a mite, a significantly higher percentage of dead flies of this species was recorded than in the control.   .B. mali females can not only be transported by females of the flightless forms of D. melanogaster and D. hydei but can also feed on them during transport and negatively affect their viability. This indicates an ectoparasitic relationship between this mite and the flies of the Drosophillidae family, as previously suggested by Lehtinen and Aspi [D. melanogaster and D. hydei showed defensive reactions. However, the species of fly had a significant impact on the effectiveness of the mite’s attack, as well as on its subsequent attachment to the fly’s body.This study showed that and Aspi , as welland Aspi . In our B. mali females took every opportunity to get onto the body of the flies and start feeding. This behavior was innate, as the tested mites had no previous experience with fruit flies or other insects. The whole process of embarkment on the host, however, could take B. mali females up to several dozen minutes, which might have been partly due to intense defense by the flies.In the observational chambers, hungry B. mali females favored the capture of flies by the tarsus or wing. In their general plan, they are typical of most free-living mesostigmatic predatory mites, i.e., they are robust and chelate-dentate with retractable movable and fixed digits. In the Mesostigmata, these are used for a variety of purposes, including capture, penetration of the victim’s cuticle, and delivery of salivary enzymes that facilitate prey liquefaction [B. mali have relatively strongly curved ends, the movable digit has two teeth, and the fixed digit has one tooth [B. mali, there seems to be a kind of compromise between its polyphagy and the necessity of (facultative) transportation on an insect body. Other ectoparasites of drosophilid fruit flies with a similar structure of chelate-dentate chelicera may face this problem, such as M. subbadius [Proctolealeps regalis DeLeon [Blattisocius, namely B. patagorium and noctuid moths, led to the development of an elongated edentate movable digit with a simultaneous reduction of the immobile digit. Individuals of this species not only fed on these insects but also mated, and the offspring could reach adulthood on the original host [B. patagiorium are short and robust. However, as shown by Treat [Undoubtedly, the morphology of the chelicera of efaction ,51. The ne tooth ,52. Theyubbadius  or Procts DeLeon . Piercins DeLeon ,55. It inal host . Interesby Treat , the larD. hydei flies were undoubtedly more ‘difficult’ hosts for B. mali. A greater percentage of female mites did not attack D. hydei at all, or their attack ended in failure. Perez-Leanos et al. [M. subbadius clearly preferred representatives of the subgenus Drosophila and repleta group (D. hydei) as opposed to the subgenus Sophophora (D. simulans Sturtevant), both in the field and in choice and no-choice experiments on immobilized (which were incapable of defending themselves) flies. It cannot be ruled out that B. mali may also have a preference for D. melanogaster in contrast to D. hydei. However, this requires further research. Undoubtedly, however, D. hydei defended itself more effectively while removing the mites from the tarsi. As a result, within the first hour of observation, more than 50% of the attacks ended in the failure of mite attachment to the tarsi, and of all B. mali females tested, only 1/3 finally entered the body and drilled their chelicerae into the abdomen, cervix, or thorax.Our research showed that the species of fruit fly had a significant impact on the effectiveness of the mite’s attack, as well as on the way it entered and where it located on the body of the flies. Although both species of flies used similar tactics to throw off mites , the s et al.  point toD. nigrospiracula Patterson and Wheeler [M. subbadius, the flies flew up into the air, and they could also approach mites and appeared to exhibit reflex behavior in the form of sudden, brisk movements away from a mite. It should be mentioned that D. hydei is larger than D. melanogaster [D. hydei could be more resistant to stress than D. melanogaster, which may also have resulted in its higher persistence in defense. In our study, the fruit flies were held in quite stressful conditions; apart from the exposure to the presence of mites, they were kept in a confined space, without the opportunity to escape and without food. Flying forms of D. hydei appear to have a higher tolerance to some kinds of stress, such as starvation and heat knockdown temperatures [D. melanogaster may be more sensitive to stress in comparison to normal flies. Our 24 h observations appear to indicate differences in stress tolerance between the flightless forms of the two species. While the mortality of D. hydei in the control chamber (without mites) was 20%, in D. melanogaster, it was close to 80%.Drosophilid fruit flies’ defenses against ectoparasitic mites have been observed in  Wheeler . Their bnogaster ,58,59, aeratures ,61. As seratures , some stB. mali females usually attempted to get onto flies when the insects stopped walking. Interestingly, they started to attack D. hydei after a longer time than D. melanogaster. It is likely that D. hydei flies were simply more mobile, and therefore, the mites had to wait longer for a convenient moment to attack. Drosophila melanogaster more frequently pressed into the narrow end of the chamber, where she also curled her legs. This behavior clearly favored the mites’ embarkment, as it was from the tarsus of a curled leg that B. mali females most frequently got onto the fly’s body. On the other hand, mites more frequently used the point of contact of the wall with the other fly’s body parts in order to enter D. hydei than D. melanogaster. One explanation for this could be that D. hydei flies are larger than D. melanogaster, and the contact of their bodies with the wall could have occurred more frequently. This, in turn, could be eagerly used by a mite. However, since the attacks directed at D. hydei tarsi often ended in defeat, B. mali females could seek ‘safer’ ways to enter the body of this fly species. This can be exemplified by the parasitoid Trichogramma brassicae Bezdenko, which is occasionally phoretic on white cabbage butterflies [Our study showed that terflies . It avoiB. mali females. Indeed, these places have a flexible, intersegmental cuticle and were relatively safe, i.e., out of the reach of the tarsi of all three pairs of legs during the fly’s grooming.Mites transported by insects often show a preference for attachment sites on the host body. This preference may depend on the species and sex of the host . It may D. melanogaster. On D. hydei, the mite chose both the sites on the cervix and close to coxa III, as well as those on the thorax and abdomen. This was probably due to the fact that during the first hour, some B. mali females failed to attach to the D. hydei tarsus (from which they most often moved to the preferred places on the fruit flies) or even if they managed to do so, they had not started climbing the leg yet. Others got onto D. hydei from the contact of the chamber wall with other parts of the fly’s body and drilled themselves into less preferred sites on the fly’s thorax or abdomen. During the following hours, however, some mites finally reached the preferred places, which may have been the result of a gradual weakening of D. hydei’s defenses and/or the mites’ gain in experience. Interestingly, the sites which were preferred by B. mali in our study differ from those observed by Lehtinen and Aspi [B. mali specimens, mostly deutonymphs, were located under the fruit fly head, as in our study, but with the chelicerae stuck between the legs of the first pair, not in the cervix. Kerezsi et al. [B. mali females on Ph. semivirgo; two specimens on fly females were in a similar position as that described by Lehtinen and Aspi 1992 [B. mali attached to the coxa III. Interestingly, both the cervix and coxa III were the preferred sites of M. subbadius in field studies on D. nigrospiracula and 13 other species of drosophilid fruit flies [B. mali females depended on which D. melanogaster leg had been previously attacked. When mites attacked the tarsus I, they climbed the leg and most often attached themselves to the fly’s cervix. On the other hand, those attacking tarsus III located themselves at the coxa of these legs. Undoubtedly, this shortened the time of wandering about on the fly’s body and enabled the mite to get to each of these places as soon as possible.The mite’s preference for the attachment to the fly’s cervix or at coxa III was already detected in the first hour of our experiment, but only with and Aspi  and Kereand Aspi . It shouand Aspi , the locand Aspi , four Dri et al.  reportedspi 1992 , and theit flies . Our resM. subbadius and radiolabeled D. nigrospiracula. Moreover, in the representative of the genus Blattisocius, B. patagorium, the behavior of the mite and scars on the body of some noctuid moth species suggested that this mite fed on its hosts during transportation [B. mali females was similar to that noted during their feeding on eggs and first-stage larvae of D. melanogaster and D. hydei, as well as various stages of the mold mite T. putrescentiae .As our study has shown, after drilling with their chelicerae into the body of flies, female mites, initially completely flat, gradually filled their idiosomas, most likely by sucking out hemolymph and other tissues dissolved in saliva. This feeding by predatory mites on the drosophilids carrying them was confirmed by Polak 1996  using M.ortation . AlthougB. mali individuals that attached to the tarsus or wing, as well as the individuals that had no interest in the fruit flies during the experiment, remained flat. This indicates that attack sites such as the fly’s tarsus or wing, while used by mites for attachment, are not suitable for feeding. This does not exclude, however, the possibility that the mite could feed on the tarsus of smaller prey that does not defend as vigorously as the fruit flies and has a much thinner cuticle as well. Examples are phytoseiid mites frequently attacking and then feeding on the tarsus and tibia of female spider mites [Only the er mites .B. mali. Several times, we observed that the mite, instead of getting off the freshly dead D. hydei or D. melanogaster, remained on the fly during subsequent hours with its chelicerae inserted in the same place on the insect body. Many species of mesostigmatic predators appear to be facultative scavengers, especially on freshly dead victims [Blattisocius, namely in B. keegani [Amyelois transitella Walker to the moth’s eggs increased the predator’s fertility. In our study, even if a mite remained on the dead fly for many hours, we did not observe any changes in the filling up of its idiosoma. On the contrary, there was often a decrease in the filling up of the mite body. This may mean that the content of hemolymph in the dead flies gradually decreased, which, in turn, did not allow B. mali females full engorgement. Perhaps the mite could also feed a bit on a different type of the fly’s tissue through preoral digestion. This, however, requires further detailed research.Our study also suggests possible scavenging in  victims . Scaveng keegani . InteresB. mali on the viability of female fruit flies. In D. hydei, after 24 h in the chambers with a mite, the mortality of females was twice as high as that in the combination without a mite. In D. melanogaster, a similar analysis was not possible due to the high mortality of flies already in the first hour of the test, both in combination with a mite and in the control group. The increased mortality of D. hydei could have resulted not only from B. mali feeding but also from energy expenditure incurred by the flies during defense against a mite and during attempts to remove it or due to stress . As the experiments by Luong et al. [M. muscadomesticae (Scopoli) mites, without their attachment to a D. hydei body, can significantly increase the energy expenditure of the fly, as manifested by its elevated CO2 production. A similar effect was found in D. nigrospiracula exposed to the presence of M. subbadius [M. muscaedomesticae mite significantly reduced the survival rate of the housefly. In turn, the presence of M. subbadius on the body of D. hydei and D. nigrospiracula significantly shortened the duration of their flight, limiting the possibility of their dispersal [D. nigrospiracula but also reduces the fertility of the females and males of this fly [Our experiments revealed the significant effect of the presence of g et al.  showed, ubbadius . The negubbadius , the parispersal . The mitthis fly ,70.B. mali can act as an ectoparasite in relation to drosophilids. Not only can it feed on them during transport, but it probably also affects their survival and feeds on dead fruit flies. However, since we used the flightless forms of D. melanogaster and D. hydei, further research is needed on the wild forms of these species, both in the laboratory and in the field.In summary, our research has shown that


Agaricus is a genus of fungi in the family Agaricaceae, with several highly priced edible and medicinal species. Here we describe Agaricus macrochlamys, a new species, in A. sect. Arvenses, sympatric and morphologically cryptic with the edible and medicinally cultivated mushroom, A. subrufescens. Phylogenetic analyses showed that A. macrochlamys is closely related to A. subrufescens, and that A. fiardii is a new synonym of A. subrufescens. Despite being morphologically cryptic, A. macrochlamys can be distinguished from A. subrufescens by several ITS and tef1α species-specific markers and a 4-bp insertion in the tef1α sequence. Furthermore, A. subrufescens is a cosmopolitan species, while A. macrochlamys distribution is so far restricted to Mexico, the Dominican Republic, and the United States. Agaricus L.  includes saprotrophic mushrooms of medicinal, nutraceutical and nutritional interest. The genus has a worldwide distribution comprising around 580 species  @ 202; ttggaAtgtgg @ 502; ctt[-]tGyggkg @ 528. No specimens having allelic ITS sequences characteristic of both A. subrufescens and A. macrochlamys are known, strongly implying that they are reproductively isolated, as well as being phylogenetically distinct.However, A. subrufescens; the remaining samples from Mexico (10), the Dominican Republic (6) and the USA (1) belong to the new species A. macrochlamys. In addition, A. fiardii is now considered a new synonym of A. subrufescens, based on the molecular phylogenetic analyses presented in this paper. The molecular data from twenty-four samples that were morphologically highly similar were identified as follows: two and five samples from Mexico and the Dominican Republic, respectively, belong to A. macrochlamys formed a full-supported sister clade to A. subrufescens. Because no significant morphological differences were observed between the two species, the molecular data remain essential for species identification. Indeed, in comparison with A. subrufescens, the new species has five distinguishing ITS nucleotide substitutions, of which four are unique within the A. sect. Arvenses. This level of genetic divergence is higher than is generally observed between the closely related species in the genus [According to our combined phylogenetic analyses, the new species he genus . Agaricus macrochlamys remains to be studied in detail. Its geographical range in Mexico and the Dominican Republic seems to be restricted to cloud forests . However, the ARB1353 sample from Georgia, USA, was collected in an area with subtropical weather. The life cycles of A. macrochlamys need to be compared with those of A. subrufescens, which is known to be amphithallic  and versatile [The biology of ersatile ,43. Othe