Assessment of duration of immunity in crossbred cattle immunized with glycoproteins isolated from Hyalomma anatolicum anatolicum and Boophilus microplus
To develop immunoprophylactic measures against multi-tick infestation, two glycoproteins of 34 and 29 kDa were isolated from the larvae of Hyalomma anatolicum anatolicum and Boophilus microplus, respectively, and assessed for their efficacy against experimental challenge infestations. The synergistic effect of the antigens in the presence of incomplete Freund’s adjuvant was found to confer protection (DT%) in animals against 56.48% of larvae and 52% of adults of H. a. anatolicum, while the effect was 40% against adults of B. microplus. The efficacy (E%) of the antigens in combination against larvae and adults of H. a. anatolicum was calculated as 70% and 64.3%, respectively, and 63% against adults of B. microplus. A direct correlation between anti-glycoprotein antibody response and protection against infestation was observed. Western blot analysis detected specific antigen in the sera of animals of group A. The antigens in combination with incomplete Freund’s adjuvant could protect animals from H. a. anatolicum and B. microplus infestations for at least 30 weeks. The possibility of employing the vaccination strategy in Indian conditions is discussed.

title: Control of ticks of ruminants, with special emphasis on livestock farming systems in India: present and future possibilities for integrated control—a review
abstract: India is predominantly an agricultural country with about 70% of her population dependent on income from agriculture. Although India accounts for a significant share of world’s livestock resources, livestock production is greatly affected by ticks and tick-borne diseases (TTBDs). Therefore, India represents a particularly interesting scenario for the study of TTBDs. Herein, we review the problems and opportunities for the integrated control of ticks of ruminants with special emphasis on livestock farming systems in India. Developments discussed in the review in the area of tick vaccines and other tick control measures should have an impact on the future of Indian livestock production.

title: Laboratory rearing of Theileria annulata-free Hyalomma anatolicum anatolicum ticks
abstract: Hyalomma anatolicum anatolicum is a three-host tick which transmits Theileria annulata infection in Indian cattle. Laboratory rearing of ixodid ticks is an essential requirement of any laboratory engaged with research on ticks and tick borne diseases. The Entomology laboratory of Indian Veterinary Research Institute is fully equipped with all the facilities and skilled manpower to maintain a homogenous H. a. anatolicum population throughout the year. The continuous supply of eggs, larvae and adults of H. a. anatolicum is maintained to meet out the demand of different experiments viz., preparation of tick antigens for immunization of animals, experimental challenge, isolation of genomic DNA and RNA. Maintenance of a H. a. anatolicum colony free of T. annulata infection is imperative for the experimental challenge infestation on cross-bred (Bos indicus × B. taurus) calves, in order to prevent the transmission of T. annulata infection to the experimental animals. A system has been developed in the laboratory in which the larvae of H. a. anatolicum were fed on New Zealand white rabbits and the dropped fed nymphs molted to adults are fed on cross-bred calves free of T. annulata infection. This synthetic cycle prevents the transstadial transmission of T. annulata as the rabbits are unsusceptible to T. annulata infection and only the adults were fed on cross-bred animals. Moreover, absence of transovarial transmission of T. annulata prevents the chance of carry over infection to experimental animals in the next cycle.

title: Comparative diagnostic potentiality of ELISA and dot-ELISA in prepatent diagnosis of experimental Fasciola gigantica infection in cattle.
abstract: A glycoprotein (27 kDa) was isolated from crude somatic antigen of Fasciola gigantica by two steps affinity chromatography and was used in early detection of experimental fasciolosis in cattle by indirect ELISA and in dot-ELISA formats. Although, anti-27 kDa antibodies could be detected after 3 weeks post infection (WPI) by dot - ELISA which was one week later than indirect ELISA. The test, dot-ELISA, was more convenient in field application. By the test (dot-ELISA) the infection could be equally detected in animals infected with 100, 200 and 300 metacercariae of F. gigantica with high sensitivity. Further, the antigen (27 kDa) was not found to react with goat sera infected with Paramphistomum epiclitum, which are giving strong reaction to homologous immature and mature fluke antigens of P. epiclitum.

title: Comparative immunopotentiating properties of saponin and incomplete Freund's adjuvant coupled to affinity purified larval antigen of Hyalomma anatolicum anatolicum.
abstract: Comparative immunostimulatory properties of saponin and incomplete Freund's adjuvant (IFA) were studied using affinity purified 39 kDa larval antigen of Hyalomma anatolicum anatolicum. Significant antibody response to 39 kDa antigen was detected in the sera of rabbits immunized with both 39 kDa plus saponin and 39 kDa plus IFA in comparison to their corresponding control animals. Insignificant differences were noted in the antibody response between the animals of two immunized groups. Upon challenge, the animals immunized with 39 kDa plus IFA rejected 76.2 +/- 9.7% of larvae and 45.8 +/- 4.1% of adults while in group of animals injected with 39 kDa plus saponin rejected 80.9 +/- 11.2% of larvae and 47.2 +/- 5.7% of adults.

title: In vitro and in vivo efficacy of Acorus calamus extract against Rhipicephalus (Boophilus) microplus
abstract: To develop a environment friendly control measure against cattle tick, Rhipicephalus (Boophilus) microplus, medicinally important plants were identified and extracts were prepared. Twelve 95% ethanolic, thirteen 50% hydroethanolic and nine hot water extracts were prepared and tested against laboratory reared homogenous colony of R. (B.) microplus. Amongst the 34 extracts, 26 extracts showed no mortality within 72 h of application while 12.0 ± 4.9% to 35.0 ± 9.6% mortality of treated ticks was recorded in other extracts. Of the effective extracts, the extract prepared from rhizome of Acorus calamus proved highly efficacious and 100% final mortality within 14 DPT was recorded. The LC85 value of the extract was determined as 11.26. In vivo experiments confirmed the efficacy of the extract up to 42%, and repeat application was required after 7 DPT. The extract was found safe and no reaction was observed when animals were treated with 50% of the concentration, which was five times of the concentration used for in vivo studies. The possibility of using the extract for the control of cattle tick is discussed.

title: Diagnostic accuracy in retinoblastoma.
abstract: This study was done to ascertain the accuracy of diagnosis and possible risk of misdiagnosis in suspected retinoblastoma cases by different pre-operative diagnostic procedures like fundoscopy, ultrasonography, computerised tomography, and aqueous cytology study. Twenty-four eyes excised with diagnosis of suspected retinoblastoma were included in the study. Final diagnosis was reached by histopathological examination of excised eyes and compared with pre-operative diagnosis by different diagnostic procedures and accuracy of each procedure was calculated. Twenty-two cases were finally diagnosed as retinoblastoma and 2 cases (8.33%) as other than retinoblastoma. Computerised tomography and aqueous humour cytology were two most specific (100%) tests. Computerised tomography was the most sensitive (95.4%) test. Aqueous cytology was poorly sensitive (9.09%). Both ultrasonography and fundoscopy demonstrated sensitivity over 90% but with low specificity (50%). When test results of fundoscopy and ultrasonography were combined, specificity of this combination was 100% and sensitivity was 95.4%, which match exactly the values of computerised tomography. Combination of fundoscopy and ultrasonography has accuracy as high as that of computerised tomography. In patients with clear media this combination can be used as an effective alternative to computerised tomography to reach a diagnosis with comparable accuracy.

title: The preparation, germination properties and stability of superdormant spores of Bacillus cereus.
abstract: To determine yields, germination and stability of superdormant Bacillus cereus spores.



Superdormant B. cereus spores were isolated by germination with high concentrations of inosine or L-alanine in 2-5% yield and did not germinate with high concentrations of either of these germinants, but germinated like starting spores with Ca-DPA, dodecylamine, L-alanine plus inosine or concentrated complete medium. Yields of superdormant spores from germinations with low inosine concentrations were higher, and these spores germinated poorly with low inosine, but relatively normally with high inosine. Yields of superdormant spores were also higher when nonheat-activated spores were germinated. Superdormant spores stored at 4 degrees C slowly recovered some germination capacity, but recovery was slowed significantly at -20 degrees C and -80 degrees C.



Factors that influence levels of superdormant B. cereus spores and the properties of such spores are similar to those in B. megaterium and B. subtilis, suggesting there are common mechanisms involved in superdormancy of Bacillus spores.



Superdormant spores are a major concern in the food industry, because the presence of such spores precludes decontamination strategies based on triggering spore germination followed by mild killing treatments. Studies of the properties of superdormant spores may suggest ways to eliminate them.

title: Regenerative capability of upper and lower jaws in the newt.
abstract: The regenerating amphibian jaw represents an important model for studying pattern formation and the mechanisms underlying regeneration of facial structures. We have studied regeneration of upper and lower jaws in the urodele amphibian, Notophthalmus viridescens, using whole mount preparations stained for bone and cartilage, scanning electron microscopy and immunocytochemistry to further characterize these regenerating systems. In addition, we have investigated whether lower jaws of adults and larvae display similar regenerative ability. Although in adult animals the original shape of both the lower and upper jaws is rather faithfully reproduced following amputation, and the teeth and oral mucosa with its specialized sensory organs fully regenerate, significant differences in the regenerative ability of the various skeletal elements are observed. In fact, only tooth-bearing skeletal elements ossify, while the other elements of the regenerated skeleton remain cartilaginous for as long as 5 months after amputation. In contrast, a regenerated lower jaw in the larva is indistinguishable from an unamputated one at the same stage of development. Interestingly, regenerating adult jaws form directly bicuspid teeth, which are the type of teeth normally found in the adult, rather than the monocuspid teeth characteristic of larval jaws, indicating that jaw regeneration is not a recapitulation of development, in that an adult jaw blastema directly regenerates an adult jaw. Finally, we have studied the expression of tissue specific markers in normal and regenerating upper and lower jaws to establish whether the blastemal cells, which will form the missing part of the jaw, express any of these markers of the differentiated state, or are undifferentiated as suggested by their morphological appearance. Under our experimental conditions, no expression of markers of the differentiated state, such as those for muscle, cartilage and glands is detectable in early regenerates. On the contrary, the mesenchymal marker 22/31, whose expression in normal jaws is restricted to dermal fibroblasts and the dental pulp, is expressed in at least a half of the blastemal cells. The significance of these observations in relation to the origin of blastemal cells in the jaw will be discussed.

title: Molecular characterization of full-length genomic segment 2 of a bovine picobirnavirus (PBV) strain: evidence for high genetic diversity with genogroup I PBVs.
abstract: We report here the molecular characterization of a bovine genogroup I picobirnavirus strain RUBV-P detected from a 1-month-old diarrhoeic calf in eastern India. Sequence comparisons and phylogenetic analysis of a short stretch of gene segment 2 of RUBV-P revealed low nucleotide identities (51.2-64.9%) with and distant genetic relatedness to other genogroup I picobirnaviruses. The complete gene segment 2 sequence of RUBV-P was obtained by the single primer amplification method with modifications. Gene segment 2 of RUBV-P was 1758 bp long, encoded a predicted protein of 554 aa and exhibited low nucleotide (58.1-58.8%) and amino acid (51.3-55.4%) identities with genogroup I human strains Hy005102 and 1-CHN-97. The 5'- and 3'-end nucleotide sequences, and the three motifs of RNA-dependent RNA polymerases of double-stranded RNA viruses, were conserved among these strains. Our findings suggested that bovine strain RUBV-P might be distinct from genogroup I picobirnaviruses of humans and other animals.

title: Full genomic analysis of a simian SA11-like G3P[2] rotavirus strain isolated from an asymptomatic infant: identification of novel VP1, VP6 and NSP4 genotypes.
abstract: We report here the full genomic analysis of a simian SA11-like G3P[2] group A rotavirus (GAR) strain, B10, isolated from an asymptomatic infant in Kenya in 1987. By nucleotide sequence identities and phylogenetic analyses, the VP7-VP4-VP2-VP3-NSP1-NSP2-NSP3-NSP5 genes of strain B10 exhibited maximum genetic relatedness to those of the different isolates of simian strain SA11, and were assigned to the G3-P[2]-C5-M5-A5-N5-T5-H5 genotypes, respectively. On the other hand, the VP1, VP6 and NSP4 genes of strain B10 did not belong to any of the established GAR genotypes, and therefore, were assigned to new genotype numbers R8, I16 and E13, respectively, by the Rotavirus Classification Working Group. These observations suggested that strain B10 might have originated from reassortment event/s involving simian SA11-like strains and GAR strains from unknown animal host species (possibly other wild animals) preceding transmission to humans. Alternatively, considering the lack of data on simian GARs, it might be also possible that the VP1, VP6 and NSP4 genes of strain B10 are those of unknown simian strains, and that strain B10 might be a typical simian strain that was directly transmitted to humans. Therefore, either hypothesis pointed towards a rare instance of possible direct transmission of GARs from an animal host (possibly a monkey or some other wild animal) to humans. This was corroborated by the presence of different species of wild animals including non-human primates, and unhygienic conditions at the sampling site. To our knowledge, the present study is the first report on the detection of a simian SA11-like G3P[2] GAR strain in humans.