bluuebunny/crossref_metadata_2025 huggingface dataset:
Serologic and Molecular Evaluation of Leishmania infantum in Cats from Central Spain
Authors: Ayllon Tania , Tesouro Miguel A. , Amusategui Inmaculada , Villaescusa Alejandra , Rodriguez‐Franco Fernando , Sainz Ángel
Published: August 2009

Infection by different Leishmania spp. in cats has been reported in many countries. In Spain, since the first Leishmania infection described in 1933, sporadic clinical cases in cats have been reported. Various serologic studies performed in other areas of Spain have shown seroprevalences ranging between 1.7 and 60%. The aim of the present study was to determine the prevalence of leishmaniasis in cats from Central Spain (Madrid), and to assess the existence of associations between Leishmania infantum infection and relevant data obtained from each cat. Two‐hundred thirty‐three cats attended at the Veterinary Teaching Hospital in Madrid between September 2005 and June 2006 were tested for L. infantum using the indirect immunofluorescent antibody (IFA) test (cutoff: 1:100) and PCR. PCR testing was performed on the samples to detect Leishmania infection, targeting the kinetoplast DNA (kDNA). Our results showed a seroprevalence of 1.29% (3/233) using IFA test. Another seven cats were also seroreactive to L. infantum one dilution under the cutoff (1:50). Considering all the seroreactive samples, the percentage of positive animals to L. infantum was 4.29%. Only one of the cats (0.43%) included in the study was PCR‐positive. Relative lymphocytosis and an increase in alanine aminotransferase (ALT) value were statistically associated with seroreactivity to L. infantum. Our results demonstrate the presence of cats seroreactive to L. infantum in Central Spain, an endemic area for this disease in dogs.
Study on Theileria lestoquardi Antigens as Potential Vaccine Candidates
Authors: Namavari Mehdi , Hosseini Mohammad H. , Seghatoleslam Atefeh , Lotfi Mohsen , Shirazi Ali , Sparagano Olivier A. E.
Published: August 2009

Theileria lestoquardi is the causative agent of malignant theileriosis of sheep and goats, causing morbidity and mortality in these animals worldwide. Western blot analysis based on T. lestoquardi schizont antigens was carried out using sera collected from Iranian sheep, which had been immunized with T. lestoquardi schizont‐infected cells. The results of Western blot analysis demonstrated that schizont‐immunized animals produced antibodies reacting with protein bands at 73, 42, 20, 14, and 12 kDa. Comparison of the results of the current Western blotting test with earlier studies of Theileria spp. revealed two immunogenic schizont proteins with molecular weights of 73 and 42 kDa shared between T. annulata and T. lestoquardi. Two other proteins with molecular weights of 14 and 12 kDa have not been previously found in other Theileria species. Our results suggest that the 73‐kDa protein could be a potential vaccine candidate and that the 14‐ and 12‐kDa proteins could be considered as diagnostic antigens.
Field Collection and Genetic Classification of Tick‐borne Rickettsiae and Rickettsiae‐like Pathogens from South Texas: Coxiella burnetii Isolated from Field‐collected Amblyomma cajennense
Authors: Sanders David M. , Parker Jill E. , Walker Wes W. , Buchholz Matt W. , Blount Keith , Kiel Johnathan L.
Published: August 2009

We are reporting the first known isolation of the Q‐fever agent Coxiella burnetii from field‐collected cayenne ticks Amblyomma cajennense in North America. Q‐fever affects a number of domestic ungulates where it can lead to abortion in sheep and goats. There is far less known about the disease's effects on wild species, primarily because of the tendency of the disease to self resolve and to provide long‐term immunity to subsequent infections. The first recovery of C. burnetii in North America was from the tick species Dermacentor andersoni. Since the original isolation C. burnetii has been recovered from five other North American tick species. The currently accepted mode for the majority of human infections is inhalation. The Centers for Disease Control and Prevention, Division of Viral and Rickettsial Diseases, Rickettsial Zoonoses Branch asserts the Q‐fever agent as requiring as few as one organism to cause disease via inhalation in susceptible humans. However, with more and more isolations from ticks, evidence linking C. burnetii and ticks is mounting. The true role of tick species as competent vectors is still unconfirmed. Preemptive field collections of possible vector arthropods, hosts, and reservoirs can provide invaluable baseline environmental data that will prove supportive in follow‐up studies and abatement efforts.
Comparison of the Molecular Structure of the TaSP Gene of Theileria annulata from Sudanese Isolates
Authors: Ali Awadia M. , Salih Diaeldin , Bakheit Mohammed , El Hussein Abdel Rahim M. , Hassan Shawgi M. , Mukhtar Maowia M. , Ahmed Jabbar S. , Seitzer Ulrike
Published: August 2009

The polymorphic region of the Theileria annulata surface protein (TaSP) was cloned and sequenced from different isolates of cattle and cell lines from different areas of Sudan. Amino acid sequence alignment revealed a high diversity showing amino acid and length polymorphism, both within and between parasite isolates. The generation of TaSP diversity may allow the evasion of host immunity by the parasite since TaSP is a highly antigenic parasite protein.
Development of an Indirect ELISA for the Diagnosis of Equine Piroplasmosis
Authors: Asenzo G. , Wilkowsky S. , Barrandeguy M. , Mesplet M. , Benitez D. , Florin‐Christensen M.
Published: August 2009

An indirect ELISA (iELISA) for the detection of specific anti‐Theileria equi antibodies in horse serum was developed. Its performance showed good concordance (K= 0.79) when compared with a competitive ELISA recommended by the World Organisation for Animal Health. Horse serum samples from two provinces located in the north and east of Argentina (Formosa and Entre Rios, respectively) were analyzed by this iELISA. A high percentage of positive horses were found in Formosa, consistent with the climatic conditions of the region that are apt for the development of tick vectors. Surprisingly, seropositive animals were also detected, although in a lower proportion, in Entre Rios, which has a temperate weather and is presumably tick free. Breeding of thoroughbred racing horses is an important economic asset of Argentina. Since equine piroplasmosis is a constraint for horse export, the epidemiological situation in different regions of the country needs to be assessed for the implementation of control measures. The iELISA developed in this work provides a convenient tool for this type of study.
Fungal Formulations to Control Rhipicephalus sanguineus Engorged Females
Authors: Reis Rosana C. S. , Fernandes Éverton K. K. , Bittencourt Vânia R. E. P.
Published: August 2009

Entomopathogenic fungi are promising alternatives to chemical acaricides in controlling tick populations; however, biological formulations are required to improve conidial performance of these pathogens in the field. This study evaluated the effect of in vitro treatment of Rhipicephalus sanguineus with conidial formulations of Beauveria bassiana (Bb) or Metarhizium anisopliae (Ma). Conidia were formulated (at 108 conidia mL−1) in (1) emulsifiable concentrate (EC), (2) polymerized cellulose gel (PCG), or both (3) emulsifiable concentrate and polymerized cellulose gel combined (EC+PCG). Engorged females were weighed and exposed to one of six treatment groups: EC+Bb, EC+Ma, PCG+Bb, PCG+Ma, EC+PCG+Bb, EC+PCG+Ma; or to control groups: C1 (untreated) or (C2) treated with Tween 80 aqueous solution, EC or PCG. After exposure, ticks were placed in petri plates and kept at 27 ± 1°C and 80% relative humidity (RH). Ticks treated with formulated conidia of entomopathogenic fungi showed significantly higher control percentage in comparison to that of control groups. The formulation of B. bassiana with EC and PCG combined (EC+PCG+Bb) caused the highest control percentage (86.79%) of R. sanguineus. This study concludes that entomopathogenic fungus conidial formulations are important to ensure effective tick control.
In Vitro Efficacy of Amitraz and Cypermethrin on Boophilus microplus from Dairy Farms in Lara State, Venezuela
Authors: Bravo Maribel , Henríquez Humberto , Coronado Alfredo
Published: August 2009

In Venezuela, the acaricides, amitraz and cypermethrin, are widely used to control tick infestations in cattle infested with Boophilus microplus. This work aims to determine the in vitro efficacy of amitraz and cypermethrin through the adult immersion test. A sample of 480 engorged females was collected from four dairy farms (180 ticks/farm) and divided in groups of 10 ticks averaging 220 mg/tick. Commercial preparation of amitraz and cypermethrin were used to prepare the working concentration of amitraz (52, 104, 208, 416 and 832 ppm), and cypermethrin (37.5, 75, 150, 300, 600 ppm). The control groups were submerged only in water. Ticks were placed in a glass lamina at 27 °C and 85% relative humidity (RH) and incubated for 18 days. The eggs masses were collected, weighed, separated in vials, and allowed to hatch at 85% RH and 27 °C for 21 days. Both the emerged larvae and eggs were quantified in each tube in order to determine the efficacy percentage and the resistance factor. The efficacy average to amitraz was 30.89%, 50.75%, 50.62%, 59.72%, and 60.53% at concentrations of 52, 104, 208, 416, and 832 ppm, respectively. The efficacy of cypermethrin was 14.3%, 11.82%, 17.25%, 20.23%, and 43.23% at concentrations of 37.5, 75, 150, 300, and 600 ppm, respectively. The amitraz resistance factor to each farm was 3.14, 1.19, 1.88, and 1.89, whereas with cypermethrin it was 8.89, 6.03, 7.12, and 18.45. Statistically significant differences between the effectiveness of different concentrations of amitraz or cypermethrin were not observed. Higher concentrations do not increase the efficacy of the products.
Molecular Analysis of Calreticulin Expressed in Salivary Glands of Rhipicephalus (Boophilus) microplus Indigenous to Thailand
Authors: Kaewhom Paitoon , Stich Roger W. , Needham Glen R. , Jittapalapong Sathaporn
Published: August 2009

The tropical cattle tick, Rhipicephalus (Boophilus) microplus, is an important ectoparasite of livestock in Thailand that causes economic losses due to the direct effects of tick feeding and by the pathogens they transmit. Intensive acaricide use has several drawbacks, which spurred efforts to develop anti‐tick vaccines. Vaccines targeting concealed antigens localized in the tick midgut result in reduced tick fecundity, but molecules localized in the tick salivary glands, which could play a role in pathogen transmission, remain largely unexplored for R. microplus. Calreticulin (CRT) is a protein found in tick salivary glands and saliva, and CRT might facilitate tick feeding and pathogen transmission through anti‐thrombotic and complement‐inhibition activities. This then suggests that CRT should be evaluated as a vaccine candidate antigen to control cattle ticks in Thailand. The objective of this work was to clone, sequence, and analyze cDNA encoding CRT from salivary glands of R. microplus indigenous to Thailand. Nucleotide sequence analysis showed an open reading frame of 1233 bp. Comparison of the amino acid sequence showed 65–99% identities to other known CRTs from Oryctolagus cuniculus, Rattus norvegicus, Homo sapiens, Bos taurus, R. sanguineus, and R. microplus. The N‐ and P‐domains of CRT were the most conserved, whereas the C‐domain was high acid and more variable. CRT primary sequences were most conserved among mammals. Further investigations are warranted to determine whether immunization with Thai B. microplus CRT can affect tick performances and experimental pathogen transmission.
Flow Cytometry to Evaluate Anaplasma marginale Parasitemia Using a Fluorescent Nucleic Acid Stain
Authors: Moretta Rosalía , Ruybal Paula , Mesplet María , Petrigh Romina , Nuñez Pablo , Gil Gonzalo , Wilkowsky Silvina , Garbossa Graciela , Farber Marisa
Published: August 2009

In this work we describe a flow cytometry–based method using SYTO16 (a DNA intercalating agent) to quantify Anaplasma marginale–infected erythrocytes in blood from bovine animals. The linearity and reproducibility of the results obtained with SYTO16 labeling followed by flow cytometry analysis make it a suitable approach for measurement of parasitemia in A. marginale infections.
Phenotype Characterization of Pelibuey Native Lambs Resistant to Haemonchus contortus
Authors: Gonzalez Jorge L. , López‐Arellano María E. , Olazaran‐Jenkins Sara , Liébano‐Hernandez Enrique , De Gives Pedro Mendoza , Vázquez‐Prats Victor , Vega‐Murillo Vicente , Calderon Rene
Published: August 2009

Specific phenotypic traits of the blood‐feeder nematode Haemonchus contortus were characterized on Pelibuey native, cross‐breed sheep in a tropical region of Mexico with background of anthelmintic problems. Data were collected on 4‐month‐old lambs (n= 63) and analyzed weekly in three stages: stage 1 (S1, not infected (0–3 weeks)); S2, first infection with 350 H. contortus infective larvae (L3) per kg of body weight by oral route (3–10 weeks); and S3, second infection with 175 H. contortus L3 per kg of body weight by oral route (10–18 weeks). Once infection took place, sheep grazed in a free‐nematode experimental paddock, and hemoncosis‐related traits, such as the number of eggs per gram of feces (epg), packed cell volume (%pvc), and eosinophils (%eo), were measured each week. Results were analyzed on specific traits and stages using SAS statistics. Statistical differences of the main phenotypic traits (P < 0.05) were observed throughout S2 on Pelibuey high (hr) and low (lr) responders, respectively: epg mean, 475 ± 340.5 and 1439 ± 243.6; %pvc mean, 31 ± 0.3 and 27 ± 0.2; and %eo mean, 2.5 ± 0.3 and 1.99 ± 0.2. In addition, lr and hr sheep grazed for 3 months on an area infected with Haemonchus, Cooperia, and Strongyloides, and the hr sheep showed a low number of epg after being exposed to multiple infections, so that no further treatment was required. These results showed that the selection of animals resistant to nematodes can be an alternative method of control against anthelmintic resistance problems in order to improve animal health and avoid a negative environmental impact.
In Silico Fasciolicide Activity of Three Experimental Compounds in Sheep
Authors: Ibarra‐Velarde Froylán , Vera‐Montenegro Yolanda , Huesca‐Guillen Alma , Cantó‐Alarcón Germinal , Alcalá‐Canto Yazmin , Marrero‐Ponce Yovani
Published: August 2009

The aim of the present study was to evaluate the fasciolicide activity of three experimental drugs, selected by an in silico system called TOMOCOMD–CARDD, in sheep. Drugs were identified by the computer system, and, after statistical selection, 24 Pelibuey sheep were infected on days 0 and 30, each with 200 metacercariae of Fasciola hepatica. When the infection reached 8 and 4 weeks of age, respectively, four groups of six animals each were formed. Group 1 received thiacetazone 150 mg/animal/p.o. Group 2 was treated with 3,5,5, trimethyloxazolidine 2,4‐dione at 450 mg/animal/p.o. G3 received guanabenz acetate at a dose of 1.5 mg/animal/p.o. G4 served as an untreated control. Monitoring of the animals was followed by individual coprological examinations and slaughter of the animals 15 days after treatment to collect and count flukes from the liver. Efficacy was measured as the reduction in the percentage of flukes of treated animals relative to untreated controls. Results indicated an efficacy of 80.0, 43.8, and 100% for 8‐week‐old flukes and 62.1, 57.9, and 100% for 4‐week‐old flukes in the three experimental groups, respectively. Even though guanabenz acetate showed a high efficacy, it was highly toxic since two animals died approximately 24 h after being treated. We conclude that further investigations should be conducted to perform computer‐aided prediction of drugs aimed to detect fasciolicide activity.
Mitochondrial DNA Origin of Indigenous Malagasy Chicken
Authors: Razafindraibe Hanta , Mobegi Victor A. , Ommeh Sheila C. , Rakotondravao  , Bjørnstad Gro , Hanotte Olivier , Jianlin Han
Published: August 2009

We report the mitochondrial DNA (mtDNA) characterization of 77 indigenous chickens (fighting and meat birds) from Madagascar, using DNA sequences of the first hypervariable segment of the D‐loop. Comparison with reference samples from the African continent and Asia revealed two mtDNA haplogroups, suggesting a dual geographic and genetic origin for the indigenous Malagasy chickens. The most common haplogroup was present in 65 individuals of the two types; it is likely of Indonesian origin. The second haplogroup was observed in 12 fighting birds and meat chickens; it could be of African continental origin and/or the result of recent introgression with commercial lines. We further studied a G/A single nucleotide polymorphism at nucleotide position 1892 bp of the coding sequence of the Mx gene that is reported to be one of the candidate susceptible/resistant genes to viral infection in chicken. Our results indicate the “susceptible” allele G is the most common with frequencies of 65% and 70% in Malagasy fighting and meat chickens, respectively. However, the allelic frequency difference between the two types of chickens is not significant (P > 0.05). These results are discussed in light of our current linguistic and archaeological knowledge on the origin of indigenous Malagasy chickens.
Tropical Veterinary Parasites at Harvard University's Museum of Comparative Zoology
Authors: Conn David Bruce
Published: August 2009

Tropical veterinary parasites have been maintained by the Museum of Comparative Zoology (MCZ) at Harvard University since the mid 1800s. Most of these are maintained by the Department of Invertebrate Zoology, but many vectors and intermediate hosts are maintained by the Departments of Entomology and Malacology. The largest collections are of avian and mammalian ticks (Acarina) that are important as both parasites and vectors. Nematodes are second in numbers, followed by cestodes, trematodes, and several minor helminth groups, crustacean parasites of fish, and protozoan parasites of various hosts. The MCZ directed or participated in several major expeditions to tropical areas around the globe in the early 1900s. Many of these expeditions focused on human parasites, but hundreds of veterinary and zoonotic parasites were also collected from these and numerous, smaller, tropical expeditions. Host sources include companion animals, livestock, laboratory species, domestic fowl, reptiles, amphibians, exotics/zoo animals, commercially important fishes, and other wildlife. Specimens are curated, either fixed whole in vials or mounted on slides as whole mounts or histopathological sections. The primary emphasis of MCZ's current work with tropical veterinary parasites is on voucher specimens from epidemiological, experimental, and clinical research.
Development of Specific Oligonucleotide Probes to Detect Vibrio Species
Authors: Molini Umberto , Thanantong Narut , Giangaspero Annunziata , Sparagano Olivier A. E.
Published: August 2009

Many species of Vibrio are responsible for diseases in marine organisms and for economic losses to the aquaculture industry. The aim of this preliminary study was to obtain species‐specific DNA zones to be used as potential probes from a phylogenetic analysis of the 23S ribosomal RNA (rRNA) gene of different Vibrio species from marine and human organisms. Species‐specific probes were identified for V. parahaemolyticus, V. fortis, V. splendidus, and for two clusters of taxonomically related species, namely V. harveyi/campbelli and V. lentus/aestuarianus. A reverse line blot assay showed that the designed probes can specifically detect the different Vibrio species, thereby proving that these probes can be used to evaluate the presence of pathogenic and nonpathogenic Vibrio species in the sea and in marine organisms to assist in the investigation of environmental risks.
Primary Midgut, Salivary Gland, and Ovary Cultures from Boophilus microplus
Authors: Mosqueda Juan , Cossío‐Bayugar Raquel , Rodríguez Elba , Falcón Alfonso , Ramos Alberto , Figueroa Julio V. , Alvarez Antonio
Published: August 2009

Primary cell cultures from different tick organs are a valuable tool for host parasite research in the study of the protozoan Babesia sp., which infects different organs of the tick. In this work we describe the generation of midgut, salivary gland, and ovary primary cell cultures from dissections of Boophilus microplus. Midguts, salivary glands, and ovaries were dissected from B. microplus ticks on different days after bovine infestation; different enzymatic disaggregating protocols were tested in the presence of proteolytic enzymes, such as trypsin and collagenase type I and II, for tissue disaggregation and primary cell culture generation. The dissected tick organs obtained 18–20 days after bovine infestation showed a major cellular differentiation and were easier to identify by cellular morphology. The enzymatic disaggregation results showed that each tissue required a different proteolytic enzyme for optimal disaggregation; collagenase type I produced the most complete disaggregation for ovaries but not for midgut or salivary glands. Collagenase type II was effective for salivary glands but performed poorly on ovaries and midgets, and typsin was effective for midguts only. The midgut and ovary primary cell cultures were maintained for 4 weeks in optimal conditions after the cells were no longer viable. The salivary gland cell cultures were viable for 8 months.
A Resolution by the Society for Tropical Veterinary Medicine in Support of “One Health”
Authors: Bokma Bob Henry , Gibbs Edward Paul Johnson , Aguirre Alfredo Alonso , Kaplan Bruce
Published: August 2009

At its ninth biennial conference, the membership of the Society for Tropical Veterinary Medicine (STVM) adopted a resolution in support of “One Health,” an initiative that promotes collaboration between veterinary and human medical professionals and allied health scientists. The resolution is published on the website of the STVM and has been shared with related veterinary medical and medical associations and societies.
Community‐based Animal Health Care
Authors: Bhandari Dilip P. , Wollen Terry S.
Published: August 2009

Livestock contribute to the livelihoods of around 70% of the world's poor. Heifer International provides high quality livestock, training, and related support to communities to assist with food security and to improve livelihoods. Heifer provides training and extension services so families can provide improved animal housing, management, breeding, nutrition, and veterinary health. Heifer conducts training programs for para‐vets or Community Animal Health Workers (CAHWs) in areas where local professional veterinary care is not available. Beneficiaries get animal health and husbandry services through trained technicians who are selected from within the community and trained in improved local practices, using basic selection guidelines. The selected community men and women get hands‐on training based on the principle of community‐based animal health care, using participatory principles in the farmers' fields. Basic tasks of CAHWs in the Heifer‐supported project are described. Heifer International is providing sustainable solutions to the well‐being of the livestock where it works.
Differential Expression of the Tick Protective Antigen Subolesin in Anaplasma marginale‐ and A. phagocytophilum‐infected Host Cells
Authors: De La Fuente José , Blouin Edmour F. , Manzano‐Roman Raúl , Naranjo Victoria , Almazán Consuelo , De La Lastra José Manuel Pérez , Zivkovic Zorica , Massung Robert F. , Jongejan Frans , Kocan Katherine M.
Published: August 2009

Subolesin was recently shown in vaccine and RNA interference (RNAi) studies to protect against tick infestations and to affect tick feeding, reproduction, and development as well as infection of host cells by Anaplasma marginale and A. phagocytophilum. Recent experiments provided evidence that infection of both tick and vertebrate host cells with these two pathogens modified gene expression. We therefore hypothesized that infection of host cells with A. marginale and A. phagocytophilum affects expression of subolesin. Subolesin mRNA levels were determined by real‐time reverse transcriptase (RT)‐PCR in uninfected and A. marginale‐infected Dermacentor variabilis guts and salivary glands and IDE8‐cultured tick cells and in uninfected and A. phagocytophilum‐infected Ixodes scapularis nymphs, ISE6‐cultured tick cells, and the human cell line HL‐60. In addition, the effect of subolesin on Anaplasma spp. infection/multiplication was characterized by RNAi in tick tissues and/or cultured tick and human cells. These experiments presented evidence of differential expression of subolesin in A. marginale‐ and A. phagocytophilum‐infected cells. Subolesin was differentially expressed in A. marginale‐infected ticks in a tissue‐specific manner in which mRNA levels increased in response to A. marginale infection in tick salivary gland cells but not in the gut cells. Subolesin knockdown by RNAi reduced Anaplasma infection/multiplication only in cells in which infection increased subolesin expression, i.e., in A. marginale‐infected D. variabilis salivary glands and IDE8 cells. The results reported herein further support the role of subolesin in Anaplasma–host interactions and suggest a putative role of subolesin in vaccines for the control of pathogen infection/multiplication in ticks.
Immunological Control of the Poultry Red Mite
Authors: Arkle Sam , Harrington David , Kaiser Pete , Rothwell Lisa , De Luna Carlos , George David , Guy Jonathan , Sparagano Olivier A. E.
Published: August 2009

In the current study whole poultry red mite antigens were extracted and birds were immunized subcutaneously with either antigen in adjuvant (antigen group) or PBS in adjuvant (control group). Immune responses of birds following immunization were investigated by ELISA and Western blotting, while vaccine efficacy was assessed by feeding of red mites on birds. Immunized birds showed a significant (P < 0.05) increase in IgY titers after immunization compared to controls, while immunoglobulin A (IgA) and IgM did not change significantly. However, the antigen group had a generally higher increase in all immunoglobulin titers compared to the controls. Western blotting identified a number of protein bands at different molecular weights, although these were not different between treatments. PCR analysis of whole mite protein identified bacterial DNA that might have confounded immunological data. In addition, there was a trend toward reduced survival rate of red mites feeding on antigen‐immunized birds, but the difference was not statistically significant compared to controls. This study demonstrates the potential for somatic red mite antigens to stimulate an antibody‐mediated immune response, although this response did not confer protection to birds.
“Pathogeno‐Proteomics”
Authors: Holzmuller Philippe , Grébaut Pascal , Brizard Jean‐Paul , Berthier David , Chantal Isabelle , Bossard Géraldine , Bucheton Bruno , Vezilier Frederic , Chuchana Paul , Bras‐Gonçalves Rachel , Lemesre Jean‐Loup , Vincendeau Philippe , Cuny Gérard , Frutos Roger , Biron David G.
Published: August 2009

Many scientists working on pathogens (viruses, bacteria, fungi, parasites) are betting heavily on data generated by longitudinal genomic–transcriptomic–proteomic studies to explain biochemical host–vector–pathogen interactions and thus to contribute to disease control. Availability of genome sequences of various organisms, from viruses to complex metazoans, led to the discovery of the functions of the genes themselves. The postgenomic era stimulated the development of proteomic and bioinformatics tools to identify the locations, functions, and interactions of the gene products in tissues and/or cells of living organisms. Because of the diversity of available methods and the level of integration they promote, proteomics tools are potentially able to resolve interesting issues specific not only to host–vector–pathogen interactions in cell immunobiology, but also to ecology and evolution, population biology, and adaptive processes. These new analytical tools, as all new tools, contain pitfalls directly related to experimental design, statistical treatment, and protein identification. Nevertheless, they offer the potency of building large protein–protein interaction networks for in silico analysis of novel biological entities named “interactomes,” a way of modeling host–vector–pathogen interactions to define new interference strategies.
The Effect of Habitat Fragmentation and Species Diversity Loss on Hantavirus Prevalence in Panama
Authors: Suzán Gerardo , Marcé Erika , Giermakowski J. Tomasz , Armién Blas , Pascale Juan , Mills James , Ceballos Gerardo , Gómez Andres , Aguirre A. Alonso , Salazar‐Bravo Jorge , Armién Anibal , Parmenter Robert , Yates Terry
Published: August 2009

Habitat fragmentation and diversity loss due to increased conversion of natural habitats to agricultural uses influence the distribution and abundance of wildlife species and thus may change the ecology of pathogen transmission. We used hantaviruses in Panama as a research model to determine whether anthropogenic environmental change is associated with changes in the dynamics of viral transmission. Specifically, we wanted to determine whether hantavirus infection was correlated with spatial attributes of the landscape at both large and small scales or whether these changes are mediated by changes in community composition. When analyzed at coarse spatial scales, hantavirus reservoirs were more commonly found in disturbed habitats and edge habitats than in forested areas. At local scales, reservoir species dominance was significantly correlated with the slope of the terrain. To evaluate the effect of small‐mammal diversity loss on infection dynamics, we implemented an experiment with selective species removal at experimental sites. Seroprevalence of hantavirus was higher in the community of small mammals and increased through time in the experimental sites. The higher seroprevalence in experimental plots suggests that greater diversity likely reduces encounter rates between infected and susceptible hosts. Our studies suggest that habitat loss and fragmentation and species diversity loss are altering hantavirus infection dynamics in Panama. Our work represents a multidisciplinary approach toward disease research that includes biodiversity concerns such as environmental change and degradation, human settlement patterns, and the ecology of host and nonhost species, work that may be especially important in tropical countries.
Recent Discoveries of New Hantaviruses Widen Their Range and Question Their Origins
Authors: Henttonen Heikki , Buchy Philippe , Suputtamongkol Yupin , Jittapalapong Sathaporn , Herbreteau Vincent , Laakkonen Juha , Chaval Yannick , Galan Maxime , Dobigny Gauthier , Charbonnel Nathalie , Michaux Johan , Cosson Jean‐François , Morand Serge , Hugot Jean‐Pierre
Published: August 2009

Hantaviruses belong to the Bunyaviridae family. While usually hosted by wild mammals, they are potentially pathogenic for humans, and several serologically distinct groups associated with different syndromes have been identified. Yet, investigations have mostly been conducted where human infections by hantaviruses constitute a real and well‐identified public health problem, i.e., the holarctic and neotropical areas. Some hantaviruses have also been described from a Suncus murinus in India and a Bandicota indica in Thailand. In addition, recent investigations in Cambodia revealed new Hantavirus types. More recently, two new Hantavirus species were described: Sangassou from a Hylomyscus simus, and Tanganya from a Crocidura theresae, both from Africa (Guinea), thus strongly questioning the current views about geographic range, evolution, and epidemiology of hantaviruses. In such a framework, we have conducted a survey of Hantavirus diversity in Southeast Asia which allows us to isolate the Thailand virus and address questions about the taxonomy of their rodent hosts. Here we present a molecular analysis of representatives of all currently known Hantavirus species, thus allowing the comparison between the newly described ones with a large range sample of rodent hantaviruses. Our results clearly point to the presence of a particular lineage of hantaviruses in Southeast Asia. It also strongly suggests that new viruses, additional mammalian hosts and different related syndromes in humans are likely to be discovered in the near future, particularly in Southeast Asia and in Africa, where Muridae rodents are highly diversified. Furthermore, additional work is also urgently needed to investigate the hantaviruses associated with Crociduridae and Soricidae.
Characterization of Anaplasma Infections in Sicily, Italy
Authors: Torina A. , Alongi A. , Naranjo V. , Scimeca S. , Nicosia S. , Di Marco V. , Caracappa S. , Kocan K. M. , De La Fuente J.
Published: August 2009

This study aimed to characterize infection with Anaplasma marginale, A. phagocytophilum, A. ovis, and A. platys in humans, animals, and ticks in Sicily, Italy, during 2003–2006. Serologic (competitive ELISA [cELISA]) and indirect immunofluorescence antibody [IFA]; N= 1990) and DNA (polymerase chain reaction [PCR]; N= 2788) tests were conducted on horse, donkey, cattle, sheep, goat, pig, dog, cat, roe deer, wild boar, human, and tick samples. The results reported herein suggested that in Sicily cattle are a major reservoir for A. marginale, dogs for A. platys, and sheep and goats for A. ovis. Domestic animals, such as cattle, horses, donkeys, sheep, dogs, and cats, may serve as reservoir for A. phagocytophilum, but different strains may infect ruminants and humans. All Anaplasma spp. characterized in Sicily had some distinctive genotypes for this region. Low genetic diversity was observed in A. ovis and A. platys, whereas A. marginale and A. phagocytophilum strains showed high genetic diversity. These results expanded our knowledge about the prevalence of Anaplasma spp. in Sicily and provided information to understand the epidemiology of these infections and implement measures to diagnose, treat, and control transmission to humans and animals in this region.
Anaplasma marginale Yucatan (Mexico) Strain
Authors: Camarillo Sergio D. Rodríguez , Ortiz Miguel Ángel García , Ramírez Edmundo E. Rojas , Alarcón Germinal J. Cantó , De La Torre Jesús F. Preciado , Cruz Rodrigo Rosario , Aragón Juan A. Ramos , Torres Ramón Aboytes
Published: August 2009

Anaplasma marginale Yucatan strain was found to have low virulence in cattle. We studied the virulence of this isolate by experimental inoculation of 113 susceptible cattle at increasing doses, after which only one animal required treatment for clinical disease. Subsequently, 104 cattle received a live vaccine of this strain by inoculation, which induced immunoprotection after heterologous challenged exposure with a different A. marginale isolate. In this study 14% of the immunized cattle required treatment as compared with the control nonimmunized cattle, in which 56% required treatment. The A. marginale vaccine strains used for the immunization studies had MSP1a variable regions that were different from those used for the challenge exposure.
Pathologic Evidence of Ehrlichiosis in Calves Inoculated with Ehrlichia chaffeensis
Authors: Delos Santos Jose R. C. , Oglesbee Michael , Rikihisa Yasuko , Stich R. W.
Published: August 2009

An immunocompetent animal disease model based on infection with Ehrlichia chaffeensis would facilitate research toward understanding mechanisms responsible for the broad range of clinical signs associated with human monocytic ehrlichiosis (HME). The adaptability of this model for the experimental feeding of tick species and stages and for testing therapies comparable to those for human diseases are additional advantages of large animal models. Herein, we summarize pathology reports for calves that developed fatal disease after experimental inoculation with E. chaffeensis. Elevated liver enzyme levels and lung pathology among these calves corroborated earlier reports of severe HME. Thus, an experimental disease model based on infection of outbred immunocompetent hosts with E. chaffeensis could be within our grasp for the first time.
Phylogenetic Analysis of MexicanBabesia bovisIsolates UsingmsaandssrRNAGene Sequences
Authors: Genis Alma D. , Mosqueda Juan J. , Borgonio Verónica M. , Falcón Alfonso , Alvarez Antonio , Camacho Minerva , De Lourdes Muñoz Maria , Figueroa Julio V.
Published: August 2009

Variable merozoite surface antigens ofBabesia bovisare exposed glycoproteins having a role in erythrocyte invasion. Members of this gene family includemsa‐1andmsa‐2(msa‐2c,msa‐2a1,msa‐2a2, andmsa‐2b).Small subunit ribosomal(ssr)RNAgene is subject to evolutive pressure and has been used in phylogenetic studies. To determine the phylogenetic relationship amongB. bovisMexican isolates using different genetic markers, PCR amplicons, corresponding tomsa‐1,msa‐2c,msa‐2b, andssrRNAgenes, were cloned and plasmids carrying the corresponding inserts were sequenced. Comparative analysis of nucleotide and deduced amino acid sequences revealed distinct degrees of variability and identity among the coding gene sequences obtained from 12 geographically differentB. bovisisolates and a reference strain. Overall sequence identities of 47.7%, 72.3%, 87.7%, and 94% were determined formsa‐1,msa‐2b,msa‐2c, andssrRNA, respectively. A robust phylogenetic tree was obtained withmsa‐2bsequences. The phylogenetic analysis suggests that MexicanB. bovisisolates group in clades not concordant with the Mexican geography. However, the Mexican isolates group together in an American clade separated from the Australian clade. Sequence heterogeneity inmsa‐1, msa‐2b, andmsa‐2ccoding regions of MexicanB. bovisisolates present in different geographical regions can be a result of either differential evolutive pressure or cattle movement from commercial trade.
Molecular Characterization of Babesia bovis Strains Using PCR Restriction Fragment Length Polymorphism Analysis of the msa2‐a/b Genes
Authors: Wilkowsky Silvina , Farber M. , Gil G. , Echaide I. , Mosqueda J. , Alcaraz E. , Suarez C. E. , Florin‐Christensen M.
Published: August 2009

The merozoite surface antigen‐2 (msa‐2) family of Babesia bovis is a group of variable genes that share conserved 5′ and 3′ ends and encode for membrane‐anchored glycoproteins that have been postulated as vaccine candidates. In this work, we analyzed the sequences of three of these genes (msa‐2a1, a2, and 2b) from two geographically distant strains and detected a certain degree of genotypic diversity that could be exploited to work out new molecular tools for the discrimination of B. bovis field samples. Here we describe a PCR restriction assay that was developed based on this observation and tested on several B. bovis strains and isolates. The results show a strain‐specific band pattern in geographically distant isolates, indicating the presence of differentially located BspMI restriction sites. This approach provides a simple method for the differentiation of American B. bovis strains.
Evaluation of Multinutritional Pellets Containing Duddingtonia flagrans Chlamydospore for the Control of Ovine Haemonchosis
Authors: Aguilar Juan Antonio Casillas , De Gives Pedro Mendoza , López‐Arellano María Eugenia , Hernández Enrique Liébano
Published: August 2009

This research evaluated the use of multinutritional pellets (MP) containing Duddingtonia flagrans chlamydospores as vehicles for oral administration in sheep against Haemonchus contortus. Pellets containing urea, wheat coat, soy meal, molasses, and D. flagrans chlamydospores were offered for 10 days to six nematode‐free sheep (group 1, treated). Group 2 (control), also six nematode‐free sheep, was supplied for the same period with nonfungi pellets. Feces were collected daily from each animal and deposited on water agar plates containing 500 H. contortus infective larvae. Three plates were used as three replicates per treatment, and all plates were incubated for 7 days. Microscopic observations revealed trap formation and the captured larvae. Washes from the plate content were passed through a Baermann funnel to collect noncaptured larvae and to quantify nematode reduction by fungal action. A larval reduction of 82% was found in fungal culture plates. The use of MP containing D. flagrans chlamydospores could be considered as a viable alternative control tool against sheep haemonchosis.
Lethal Activity of Two Bacillus thuringiensis Strains against Haemonchus contortus Histotropic Larvae
Authors: Linares Ismael Hernández , Arellano María Eugenia López , De Gives Pedro Mendoza , Hernández Enrique Liébano , De La Parra Alejandra Bravo
Published: August 2009

The in vitro lethal activity of two Bacillus thuringiensis strains (IB‐16 and IB‐61) against Haemonchus contortus histotropic fourth stage larvae (L4) was evaluated. Each B. thuringiensis strain was used as soluble proteins at different concentrations. In addition, the in vitro lethal dose (LD50) of each strain was determined against H. contortus L4 at different times. Data were analyzed by ANOVA and Tukey tests. Less than 50% mortality was observed at 50 and 100 μg/mL during all bioassays. More than 50% mortality was observed at 200 μg/mL at 3 and 5 days after interaction with IB‐16 and IB‐61, respectively. A 100% mortality was observed with IB‐16 on day 5. The LD50 with IB‐16 was 248 μg/mL on day 3, and IB‐61 showed an LD50 at 227 μg/mL on the day 5. Both toxins may have potential as biocontrol agents.
Amblyomma variegatum Ticks and Heartwater on Three Caribbean Islands
Authors: Vachiéry Nathalie , Jeffery Helena , Pegram Rupert , Aprelon Rosalie , Pinarello Valérie , Kandassamy Ranleen Lloyd Yane , Raliniaina Modestine , Molia Sophie , Savage Hazel , Alexander Randolph , Frebling Mathieu , Martinez Dominique , Lefrançois Thierry
Published: August 2009

Amblyomma variegatum tick infestation, tick infection by Ehrlichia ruminantium (ER), and ER genetic diversity were studied in the Caribbean Islands of Guadeloupe, Marie‐Galante, and Antigua between 2003 and 2005. Nested PCR for pCS20 was used to detect ER, while ER strains were characterized by sequencing or by restriction fragment length polymorphism (RFLP) profiles of map‐1 PCR products. In 2003 in Guadeloupe, the prevalence of tick‐infested herds was 35.6%. In Marie‐Galante 79.1% of herds in 2003 and 73.8% in 2005 were infested, while only an average of 2.2% of the herds were infected in Antigua between this same period. In Marie‐Galante, 19.1% of ticks were ER positive, and ER‐infected ticks were found in 33.3% of the herds. In Antigua only 5.8% of the ticks were ER positive. High ER tick infection rate combined with a very high level of tick infestation highlight the risk of heartwater in Marie‐Galante and Guadeloupe more than in Antigua. The three islands still represent a reservoir for tick and heartwater in the Caribbean. Nine different African and Caribbean map‐1 ER genotypes were identified. This diversity was observed even in restricted areas, and identical map‐1 genotypes were observed on all three islands. This high genetic diversity of ER strains suggests that there was a simultaneous introduction of several strains from African countries into the Caribbean region.
Immunization with Tick Salivary Gland Extracts
Authors: Jittapalapong Sathaporn , Phichitrasilp Thanmaporn , Chanphao Hathairat , Rerkamnuychoke Worawut , Stich Roger W.
Published: August 2009

Tick burdens cause direct damage to hosts and transmit several disease agents, the majority of which are secreted into feeding lesions through tick salivary glands. Reduced incidence of naturally transmitted tick‐borne diseases was recently observed among cattle immunized with tick salivary gland extracts (TSGE). The aim of this work was to compare the ultrastructure of salivary glands from Rhipicephalus (Boophilus) microplus removed from cattle that were immunized with TSGE to those from control cattle injected with saline. Partially fed female ticks that infested these cattle were collected, and their salivary glands were removed and compared with light microscopy. More extensive salivary gland damage was observed in ticks collected from cattle immunized with TSGE than those from saline controls. Salivary glands from immunized cattle had extensive necrosis in agranular c and f cells; partial necrosis in granular b, c, and e cells; and slight necrosis in a and d cells. These results indicated that host immunization with TSGE causes detrimental effects to female tick salivary glands and correlated with the reduced incidence of naturally transmitted tick‐borne diseases observed among these cattle.
Antibiotic Clearance of Ehrlichia canis from Dogs Infected by Intravenous Inoculation of Carrier Blood
Authors: Schaefer John J. , Kahn Jonathan , Needham Glen R. , Rikihisa Yasuko , Ewing S. A. , Stich R. W.
Published: August 2009

Ehrlichia canis is the etiologic agent of canine monocytic ehrlichiosis (CME) and is a useful model for zoonotic tick‐borne pathogens, many of which infect dogs. The purpose of this study was to evaluate rifampin and doxycycline regimens for clearance of E. canis infections in addition to alleviation of CME. Beagles were infected with E. canis by intravenous inoculation with carrier blood and treated with either rifampin or doxycycline after the acute phase of CME. Improved hematological values demonstrated that both treatments effectively relieved signs of the disease. Peripheral blood from all dogs became PCR negative after antibiotic treatment, suggesting that these infections were eliminated and that rifampin is an effective alternative chemotherapeutic agent for treatment of CME.
Viral Association with the Elusive Rickettsia of Viper Plague from Ghana, West Africa
Authors: Kiel Johnathan L. , Gonzalez Yvette , Parker Jill E. , Andrews Carrie , Martinez Dominique , Vachiéry Nathalie , Lefrançois Thierry
Published: August 2009

We previously reported a rickettsial heartwater‐like disease in vipers from Ghana that resembled heartwater in its gross lesions, was apparently transmitted by ticks (Aponomma and Amblyomma), and responded clinically favorably to early treatment with tetracycline. Cell culture showed consistent cytopathic effects in bovine endothelial cells, viper cells, and mouse cells, and inhibition of cytopathic effect by tetracycline in vitro. A type D retrovirus was observed in vacuoles in all infected cells. The virus and rickettsia infection was associated with transfer of cytopathic effect, regardless of cell species. Close association of virus and rickettsia may indicate a dual infection etiology of viper plague.
Mammal and Bird Species Distribution at the Woodland–Pasture Interface in Relation to the Circulation of Ticks and Pathogens
Authors: Vourc’h Gwenaël , Boyard Chloé , Barnouin Jacques
Published: August 2009

The tick vector Ixodes ricinus can be found in pastures in landscapes where pasture and woodland intermix, such as in the bocage. To identify which mammal or bird species could potentially transport ticks between woodland and pasture, we did a species inventory on four French bocage sites in 2005. Fifty‐eight species were observed, 21 of which were found in three different habitats: the middle of the pasture, the hedgerow ecotone, and the woodland. Because the bocage is a meeting point for wildlife, domestic animals, and humans, species using the three habitats could potentially be epidemiologic agents for circulation of tick vectors as well as of pathogens. Thus, the bocage could be used as a model landscape for the study of epidemiologic cycles and potential emergence of multiple‐host pathogens.