Comparison of rapid immunodiagnosis assay kit with molecular and immunopathological approaches for diagnosis of rabies in cattle

Research (Published online: 31-01-2016)

18. Comparison of rapid immunodiagnosis assay kit with molecular and immunopathological approaches for diagnosis of rabies in cattle -Ajaz Ahmad and C. K. Singh

Veterinary World, 9(1): 107-112

   doi: 10.14202/vetworld.2016.107-112

Abstract

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Aim: Presently, diagnosis of rabies is primarily based on, conventional fluorescent antibody technique (FAT), immunopathological and molecular techniques. Recently, rapid immunodiagnostic assay (RIDA) - A monoclonal antibodybased technique has been introduced for rapid diagnosis of rabies. The present investigation is envisaged to study the efficacy of RIDA kit for the diagnosis of rabies in cattle.

Materials and Methods: About 11 brain samples from cattle, clinically suspected for rabies, were screened by the FAT, Heminested reverse transcriptase polymerase chain reaction (HnRT-PCR), Immunohistochemistry (IHC), and RIDA.

Results: The sensitivity for detection of rabies from brain tissue by RIDA was 85.7% as compared to 100% by IHC as well as HnRT-PCR. The accuracy of detection of rabies by RIDA was 91.6% as compared to 100% that of IHC and HnRT-PCR, whereas specificity of RIDA was 100% like that of the IHC and HnRT-PCR.

Conclusion: Despite a comparatively low-sensitivity and accuracy of RIDA, latter can still be useful in screening a large number of field samples promptly. However, it is recommended that negative results with RIDA in cattle need to be authenticated by suitable alternative diagnostic approaches.

Keywords: cattle, diagnosis, fluorescent antibody technique, heminested reverse transcriptase, immunohistochemistry, rabies.

Anthelmintic activity of Indigofera tinctoria against gastrointestinal nematodes of sheep

Research (Published online: 31-01-2016)

17. Anthelmintic activity of Indigofera tinctoria against gastrointestinal nematodes of sheep - Ambalathaduvar Meenakshisundaram, Tirunelveli Jayagopal Harikrishnan and Thavasi Anna

Veterinary World, 9(1): 101-106

   doi: 10.14202/vetworld.2016.101-106

Abstract

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Aim: Gastrointestinal (GI) nematodes are considered as a major constraint for successful sheep production. Control of these parasites heavily relies on the use of chemical anthelmintics. Over the past decades, the development of anthelmintic resistance to various groups of anthelmintics and problem of drug residues in animal products has awakened interest in medicinal plants as an alternative source of anthelmintics. Hence, this study was undertaken to evaluate the anthelmintic efficacy of Indigofera tinctoria by scientifically validated in vitro and in vivo tests approved by the World Association for the Advancement of Veterinary Parasitology.

Materials and Methods: In vitro assays such as egg hatch assay for ovicidal and larval migration inhibition and larval development assay for larvicidal properties were used to investigate in vitro effect of extracts on strongyle egg and larvae, respectively. Fecal egg count reduction test was conducted in vivo to evaluate the therapeutic efficacy of the extracts administered orally at dose rates of 125, 250, 500 mg/kg to sheep naturally infected with mixed GI nematodes.

Results: Ethanolic extract of I. tinctoria demonstrated significant (p<0.01) inhibition on egg hatching at concentrations of 40 mg/ml and 80 mg/ml. In in vivo assay, the ethanolic extract of I. tinctoria reduced the fecal egg count ranging between 30.82% and 47.78% at various doses (125, 250 and 500 mg/kg). Although there was a slight variation, all the hematological parameters were within the normal range reported for sheep. Except for alanine transaminase, the overall mean of all the serum biochemical profile was within the normal range for sheep.

Conclusion: Based on the results obtained by in vitro and in vivo assay, the ethanolic extract of I. tinctoria possesses anthelmintic activity and could replace the chemical anthelmintics used presently.

Keywords: anthelmintic, evaluation, gastrointestinalnematodes, Indigofera tinctoria, sheep.

Potential of acute phase proteins as predictor of postpartum uterine infections during transition period and its regulatory mechanism in dairy cattle

Review (Published online: 29-01-2016)

16. Potential of acute phase proteins as predictor of postpartum uterine infections during transition period and its regulatory mechanism in dairy cattle - A. Manimaran, A. Kumaresan, S. Jeyakumar, T. K. Mohanty, V. Sejian, Narender Kumar, L. Sreela, M. Arul Prakash, P. Mooventhan, A. Anantharaj and D. N. Das

Veterinary World, 9(1): 91-100

   doi: 10.14202/vetworld.2016.91-100

Abstract

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Among the various systemic reactions against infection or injury, the acute phase response is the cascade of reaction and mostly coordinated by cytokines-mediated acute phase proteins (APPs) production. Since APPs are sensitive innate immune molecules, they are useful for early detection of inflammation in bovines and believed to be better discriminators than routine hematological parameters. Therefore, the possibility of using APPs as a diagnostic and prognostic marker of inflammation in major bovine health disorders including postpartum uterine infection has been explored by many workers. In this review, we discussed specifically importance of postpartum uterine infection, the role of energy balance in uterine infections and potential of APPs as a predictor of postpartum uterine infections during the transition period and its regulatory mechanism in dairy cattle.

Keywords: cute phase proteins, bovine uterine infections, pro-inflammatory cytokines, negative energy balance, transition period.

Recent developments in receptor tyrosine kinases targeted anticancer therapy

Review (Published online: 29-01-2016)

15. Recent developments in receptor tyrosine kinases targeted anticancer therapy - Samir H. Raval, Ratn D. Singh, Dilip V. Joshi, Hitesh B. Patel and Shailesh K. Mody

Veterinary World, 9(1): 80-90

   doi: 10.14202/vetworld.2016.80-90

Abstract

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Novel concepts and understanding of receptors lead to discoveries and optimization of many small molecules and antibodies as anti-cancerous drugs. Receptor tyrosine kinases (RTKs) are such a promising class of receptors under the investigation in past three decades. RTKs are one of the essential mediators of cell signaling mechanism for various cellular processes. Transformations such as overexpression, dysregulation, or mutations of RTKs may result into malignancy, and thus are an important target for anticancer therapy. Numerous subfamilies of RTKs, such as epidermal growth factor receptor, vascular endothelial growth factor receptor, fibroblast growth factor receptors, insulin-like growth factor receptor, and hepatocyte growth factor receptor, have been being investigated in recent years as target for anticancer therapy. The present review focuses several small molecules drugs as well as monoclonal antibodies targeting aforesaid subfamilies either approved or under investigation to treat the various cancers.

Keywords: cancer, monoclonal antibodies, small molecule drugs, receptor tyrosine kinases, targeted therapy.

Episodes of clinical mastitis and its relationship with duration of treatment and seasonality in crossbred cows maintained in organized dairy farm

Research (Published online: 21-01-2016)

14. Episodes of clinical mastitis and its relationship with duration of treatment and seasonality in crossbred cows maintained in organized dairy farm - Narender Kumar, A. Manimaran, A. Kumaresan, L. Sreela, Tapas Kumar Patbandha, Shiwani Tiwari and Subhash Chandra

Veterinary World, 9(1): 75-79

   doi: 10.14202/vetworld.2016.75-79

Abstract

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Aim: Present study aimed to evaluate the different episodes of clinical mastitis (CM) and influence of duration of treatment and seasonality on the occurrence of different episodes of CM in crossbred cows.

Materials and Methods: A total of 1194 lactation data of crossbred CM cows were collected from mastitis treatment record from 2002 to 2012. Data of CM cows were classified into types of episodes (pattern of repeated or multiple episodes occurrence) and number of episodes (magnitude of multiple cases). Types of episodes were divided as single (clinical cure by a single episode of treatment), relapse (retreatment of the same cow within 21 days), recurrence (new CM at least 21 days after treatment), and both (relapse and recurrence). The season was classified as winter (December to March), summer (April to June), rainy (July to September), and autumn (October to November). The difference between incidences of different types of CM episodes and the association between number or type of CM episodes with duration of treatment and seasons of CM occurrence were analyzed by Chi-square test.

Results: Among 1194 animals suffered with CM, 53, 16, and 18% had the single episode, relapse, and recurrence, respectively; while 13% suffered by both relapse and recurrence. We estimated the duration of treatment and found 80% of the cows treated 1-8 days, in which 65% treated for 1-4 days, while 35% cows were treated for 5-8 days. Further, 12% cows treated for 9-15 days and 7.5% cows treated >15 days. The relationship between duration of treatment and different episodes of CM revealed that 1-8 days treated cows were mostly cured by the single episode with less relapse and recurrence. In contrast, the incidences of recurrence and relapse episodes were higher in cows treated for more than 9 days. The highest incidence of relapse was noticed in winter (36%) than other seasons (10-28%), while the recurrence was less during autumn (9%) compared to other seasons (20-40%).

Conclusion: Cows those suffered by both relapse and recurrence were more susceptible to CM, and they need to be culled from farm to control the transmission of infections. Although the influence of seasonality was difficult to understand, the higher magnitude of relapse and recurrence during winter suggested the adverse effects of cold stress on treatment outcome.

Keywords: clinical mastitis, duration of treatment, episodes, recurrence, relapse, season.

Prevalence of vero toxic Escherichia coli in fecal samples of domestic as well as wild ruminants in Mathura districts and Kanpur zoo

Research (Published online: 21-01-2016)

13. Prevalence of vero toxic Escherichia coli in fecal samples of domestic as well as wild ruminants in Mathura districts and Kanpur zoo -Raghavendra Prasad Mishra, Udit Jain, Basanti Bist, Amit Kumar Verma and Ashok Kumar

Veterinary World, 9(1): 71-74

   doi: 10.14202/vetworld.2016.71-74

Abstract

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Aim: The present study was planned to reveal the prevalence of verocytotoxigenic Escherichia coli (VTEC) in fecal samples of domestic and wild ruminants in Mathura district and Kanpur zoo.

Materials and Methods: A total of 240 fecal samples comprising 60 each of cattle, buffalo, sheep and deer from Mathura districts and Kanpur zoo were screened for the presence of E. coli and VTEC genes positive by polymerase chain reaction (PCR).

Result: Out of 240 fecal samples, 212 E. coli strains were obtained. All the E. coli isolates were screened by PCR to detect virulence genes stx1, stx2, eaeA and hlyA. Of these, 25 isolates were identified as VTEC. The prevalence of VTEC in cattle, buffalo, sheep and deer was found 13.4% (8/60), 13.4% (8/60), 6.67% (4/60) and 8.33% (5/60), respectively.

Conclusion: stx1, stx2, eaeA and hlyA genes were prevalent in VTEC isolates from feces of cattle, buffalo, sheep and deer population of Mathura districts and Kanpur zoo. The presence of VTEC isolates in this region may pose a threat to public health.

Keywords: domestic and wild ruminants, feces, prevalence, polymerase chain reaction, verotoxic Escherichia coli.

Prevalence and characterization of multi-drug resistant Salmonella Enterica serovar Gallinarum biovar Pullorum and Gallinarum from chicken

Research (Published online: 20-01-2016)

12. Prevalence and characterization of multi-drug resistant Salmonella Enterica serovar Gallinarum biovar Pullorum and Gallinarum from chicken - Md. Shafiullah Parvej, K. H. M. Nazmul Hussain Nazir, M. Bahanur Rahman, Mueena Jahan, Mohammad Ferdousur Rahman Khan and Marzia Rahman

Veterinary World, 9(1): 65-70

   doi: 10.14202/vetworld.2016.65-70

Abstract

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Aim: Salmonella is an important zoonotic pathogen responsible for animal and human diseases. The aim of the present study was to determine the prevalence and stereotyping of Salmonella isolates isolated from apparently healthy poultry. Furthermore, the clonal relatedness among the isolated Salmonella serovars was assessed.

Materials and Methods: A total of 150 cloacal swab samples from apparently healthy chickens were collected, and were subjected for the isolation and identification of associated Salmonella organisms. The isolated colonies were identified and characterized on the basis of morphology, cultural characters, biochemical tests, slide agglutination test, polymerase chain reaction, and pulsed-field gel electrophoresis (PFGE). Antibiotic sensitivity patterns were also investigated using commonly used antibiotics.

Results: Of the 150 samples, 11 (7.33%) produced characteristics pink colony with black center on XLD agar medium, and all were culturally and biochemically confirmed to be Salmonella. All possessed serovar-specific gene SpeF and reacted uniformly with group D antisera, suggesting that all of the isolates were Salmonella Enterica serovar Gallinarum, biovar Pullorum and/or Gallinarum. Antimicrobial susceptibility testing revealed that 54.54% of the isolated Salmonella Enterica serovars were highly sensitive to ciprofloxacin, whereas the 81.81% isolates were resistant to amoxycillin, doxycycline, kanamycin, gentamycin, and tetracycline. Pulsed-field gel electrophoresis of the XbaI-digested genomic DNA exhibited identical banding patterns, suggesting that the multidrug resistant Salmonella Enterica serovars occurring in commercial layers are highly clonal in Bangladesh.

Conclusion: The present study was conducted to find out the prevalence of poultry Salmonella in layer chicken and to find out the clonal relationship among them. The data in this study suggest the prevalence of Salmonella Enterica, which is multidrug resistant and highly clonal for commercial layers of Bangladesh.

Keywords: polymerase chain reaction, pulsed field gel electrophoresis, Salmonella, SpeF gene.

Detection of Clostridium perfringens alpha toxin gene in lambs by loop mediated isothermal amplification

Research (Published online: 20-01-2016)

11. Detection of Clostridium perfringens alpha toxin gene in lambs by loop mediated isothermal amplification - B. Radhika, N. Vinod Kumar and D. Sreenivasulu

Veterinary World, 9(1): 60-64

   doi: 10.14202/vetworld.2016.60-64

Abstract

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Aim: The loop mediated isothermal amplification (LAMP) was standardized for rapid detection of Clostridium perfringens.

Materials and Methods: A total of 120 fecal samples were collected from enterotoxemia suspected lambs were used for screening ofC. perfringens cpa gene by LAMP. The specificity of the LAMP amplified products was tested by digesting with restriction enzyme XmnIfor alpha toxin gene.

Results: Out of 120 samples screened 112 (93.3%) samples were positive by both LAMP and polymerase chain reaction (PCR) for detection of cpa gene which indicated the equal sensitivity of both the tests. The enzyme produced single cut in 162 base pair amplified product of alpha toxin gene at 81 base pair resulting in a single band in gel electrophoresis.

Conclusion: Both LAMP and PCR for detection of cpa gene indicated the equal sensitivity of both the tests. Standardization of LAMP reaction for amplification of epsilon and beta toxin genes will help to identify the C. perfringens toxin types from the clinical samples. The test could be a suitable alternative to the PCR in detection of toxin types without the help of sophisticated machinery like thermal cycler. Considering its simplicity in operation and high sensitivity, there is the potential use of this technique in clinical diagnosis and surveillance of infectious diseases.

Keywords: Clostridium perfringens, enterotoxemia, lambs, loop mediated isothermal amplification.

Genetic structure of Mugil cephalus L. populations from the northern coast of Egypt

Research (Published online: 15-01-2016)

10. Genetic structure of Mugil cephalus L. populations from the northern coast of Egypt - Mahmoud Magdy, Mariam Gergis Eshak and Mohamed Abdel-Salam Rashed

Veterinary World, 9(1): 53-59

   doi: 10.14202/vetworld.2016.53-59

Abstract

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Aim: The gray mullet, Mugil cephalus, has been farmed in semi-intensive ponds with tilapia and carps in Egypt for years. The current study used the fluorescent amplified fragment length polymorphism (F-AFLP) technique to search for genetic differences between the populations of M. cephalus in the northern region of Egypt and to detect the gene flow between sampled locations and the homogeneity within M. cephalus genetic pool in Egypt.

Materials and Methods: To fulfill the study objectives 60 (15/location) samples were collected from four northern coast governorates of Egypt (Alexandria “sea,” Kafr El-Sheikh “farm,” Damietta “farm” and Port Said “sea”). Three replicates of bulked DNA (5 samples/replicate) for each location were successfully amplified using the standard AFLP protocol using fluorescent primers. DNA polymorphism, genetic diversity, and population structure were assessed while positive outlier loci were successfully detected among the sampled locations. Based on the geographical distribution of sampling sites, the gene flow, the genetic differentiation, and correlations to sampling locations were estimated.

Results: A total of 1890 polymorphic bands were scored for all locations, where 765, 1054, 673, and 751 polymorphic bands were scored between samples from Alexandria, Kafr El-Sheikh, Damietta and Port Said, respectively. The effective number of alleles (ne) for all bulked samples combined together was 1.42. The expected heterozygosity under Hardy–Weinberg assumption (He) for all bulked samples combined together was 0.28. Bulked samples from Damietta yielded the lowest ne (1.35) and the lowest He (0.23) when inbreeding coefficient (FIS) = 1. Bulked samples from Kafr El-Sheikh scored the highest ne (1.55) and the highest He (0.37). Bulked samples from Alexandria scored 1.40 for ne and 0.26 for He, while bulked samples from Port Said scored 1.39 for ne and 0.26 for He. The observed bulked samples formed three sub-population groups, where none is limited to a certain sampling location. A high differentiation among locations was detected, however, is not fully isolating the locations. Gene flow was 0.58. Positive outliers loci (117) were detected among the four sampled locations while weak significant correlation (r=0.15, p=0.03) was found for the distance between them.

Conclusion: Even though this species is cultivated in Egypt, the wild population is still present and by the current study a flow of its genes is still exchanged through the northern coast of Egypt. Which contribute to the cultivated populations leading to heterogeneity in its genetic pool and consequently affects the production consistency of M. cephalus in Egypt.

Keywords: fluorescent amplified fragment length polymorphism, isolation by distance, Mantel test, marine fish, Mugil cephalus, natural selection pressure, population structure.

Epidemiological study of bovine brucellosis in three senatorial zones of Bauchi State, Nigeria

Research (Published online: 15-01-2016)

9. Epidemiological study of bovine brucellosis in three senatorial zones of Bauchi State, Nigeria - S. G. Adamu, N. N. Atsanda, A. O. Tijjani, A. M. Usur, A. G Sule and I. A. Gulani

Veterinary World, 9(1): 48-52

   doi: 10.14202/vetworld.2016.48-52

Abstract

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Aim: To determine the seroepidemiological patterns of bovine brucellosis in three senatorial zones of Bauchi State, Nigeria.

Materials and Methods: Blood samples were aseptically collected from the anterior jugular vein of 336 slaughtered cattle, between September 2013 and March 2014 in three senatorial zones of Bauchi State, Nigeria. The sera obtained were screened for brucellosis using rose Bengal plate test (RBPT) and serum agglutination test (SAT) in parallel. The data generated was subjected to Chi-square and Fishers exact test analysis to establish whether there is a relationship between the breeds, sex, and location of the animals sampled.

Results: Of the 336 cattle screened, 18 (5.4%) and 13 (3.9%) were positive by RBPT and SAT, respectively. There was no statistically significant association (p>0.05) between the sex, age, and location of cattle with seropositivity of brucellosis in the state. It was concluded that brucellosis is prevalent in Bauchi State. Further study is recommended in other abattoirs and herds of cattle in Bauchi State for confirmation of the status of the disease among cattle slaughtered in the state.

Conclusion: A high seroprevalence of brucellosis among the cattle in Bauchi state indicates that the disease is endemic and cattle are one of the animals that perpetuate and sustain the disease.

Keywords: cattle, Bauchi state, brucellosis, Nigeria, seroprevalence.

Retrospective analyses of dogs found serologically positive for Ehrlichia canis in Cebu, Philippines from 2003 to 2014

Research (Published online: 13-01-2016)

8. Retrospective analyses of dogs found serologically positive for Ehrlichia canis in Cebu, Philippines from 2003 to 2014 - Adrian P. Ybañez, Rochelle Haidee D. Ybañez, Rex R. Villavelez, Honey Pearl F. Malingin, Dana Natasha M. Barrameda, Sharmaine V. Naquila and Shiella Mae B. Olimpos

Veterinary World, 9(1): 43-47

   doi: 10.14202/vetworld.2016.43-47

Abstract

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Aim: The study aimed to document the clinical and hematological observations of dogs found serologically positive for Ehrlichia canis and to identify parameters or factors that are associated with the disease with focus on the anemic and thrombocytopenic state of the infected dogs.

Materials and Methods: From 7 participating veterinary establishments, a total of 913 cases from 2003 to 2014 were initially assessed using inclusion criteria, including E. canis diagnosis by the attending veterinarian and the presence of ticks or history of infestation, thrombocytopenia, and/or anemia. From these, 438 cases that were found serologically positive for E. canis using commercial test kits were selected. Profile, clinical observations and hematological test results were obtained from the selected cases. Computations for statistical associations between the anemic and thrombocytopenic state of the infected dogs and their profile, observed clinical signs and other hematological values were performed.

Results: Most of the dogs were purebred (60.0%) and female (51.1%) and were within the age range of 1-5 years (38.4%). The mean packed cell volume (PCV), red blood cell (RBC) count, and platelet count were lower than the normal values while the absolute count of basophils were higher than normal values. Creatinine and blood urea nitrogen (BUN) appear to be elevated. The most common clinical signs observed were inappetence (41.3%), lethargy/depression (35.6%), vomiting (32.4%), fever (18.5%), paleness (8.2%), and epistaxis (6.6%). Analyses showed that there were no significant differences on the hematological values and clinical signs between thrombocytopenic and non-thrombocytopenic seropositive dogs. Moreover, very weak correlations between platelet count and RBC count, absolute lymphocyte count, and neutrophil count were found. On the other hand, only paleness (p=0.008) and epistaxis (p=0.004) were found to be significantly different between anemic and non-anemic patients. This coincided with the linear regression results where PCV (p=0.000, R=0.787, R2=0.619) was moderately correlated with the RBC count. In addition, eosinophil count was found weakly correlated.

Conclusion: E. canis infection in dogs may produce varied clinical signs that may be influenced by the thrombocytopenic and anemic states of affected animals. Complete blood counts remain important in the diagnosis of the disease, especially the platelet and RBC counts. Creatinine, BUN and alanine aminotransferase can be of value in the diagnosis of the infection. Several cases were lost to follow-up and appeared to be a challenge for handling veterinarians to monitor compliance of owners and progress of infected patients.

Keywords: Cebu, clinical signs, dog, Ehrlichia canis, hematology, Philippines.

Effect of age, sex and physiological stages on hematological indices of Banni buffalo (Bubalus bubalis)

Research (Published online: 13-01-2016)

7. Effect of age, sex and physiological stages on hematological indices of Banni buffalo (Bubalus bubalis) - Mehul D. Patel, Abdul Lateef, Hemen Das, Ajay S. Patel, Ajay G. Patel and Axay B. Joshi

Veterinary World, 9(1): 38-42

   doi: 10.14202/vetworld.2016.38-42

Abstract

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Aim: To determine the physiological baseline values for hematological indices of Banni buffalo Bubalus bubalis) as well as to assess their alteration due to age, sex and physiological stages.

Materials and Methods: A total of 42 clinically healthy Banni buffaloes were categorized into seven groups (n=6): Group I (male calves ≤1 year), Group II (bulls >1 year), Group III (female calves ≤1 year), Group IV (pregnant lactating buffaloes), Group V (non-pregnant lactating buffaloes), Group VI (pregnant dry buffaloes), and Group VII (non-pregnant dry buffaloes). Blood samples collected aseptically from all the experimental groups were analyzed employing automated hematology analyzer. The data obtained were statistically analyzed; the mean and standard deviations were calculated and set as the reference values.

Results: The erythrocytic indices viz. total erythrocytes count (TEC), hemoglobin, and packed cell volume (PCV) were significantly higher in bulls as compared to that of male calves unlike mean corpuscular volume, mean corpuscular hemoglobin (MCH), and MCH concentration. The female calves had higher TEC and PCV than the adult buffaloes irrespective of sex. The total leukocyte count (TLC) and neutrophil counts in male calves were significantly lower than the bulls unlike the eosinophil, while monocyte and basophil remained unchanged with age. The TLC, differential leukocyte count and platelet count varied non-significantly among the adult female groups at different physiological stages. However, neutrophils were found to be apparently higher in lactating buffaloes.

Conclusion: The present study would be helpful for physiological characterization of this unique buffalo breed of Gujarat. Further, data generated may be a tool for monitoring the health and prognosis as well as diagnosis of diseases.

Keywords: age, banni, erythrocytic, lactation, leukocytic indices, pregnancy, sex.

Effect of different culture systems on the production of foot and mouth disease trivalent vaccine

Research (Published online: 12-01-2016)

6. Effect of different culture systems on the production of foot and mouth disease trivalent vaccine - Amr Ismail Hassan

Veterinary World, 9(1): 32-37

   doi: 10.14202/vetworld.2016.32-37

Abstract

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Aim: This study aims to determine the effect of the stationary rawx, roller, and the suspension cell culture systems on the total virus yield infectivity and antigenicity.

Materials and Methods: Three serotypes of foot and mouth disease virus (FMDV) (serotype A, O and SAT-2) were inoculated separately into baby hamster kidney-21 cell line in rawx, roller, and suspension cultivation systems using multiplicity of infection (1:100). Samples were taken from the total virus yield from each system at 15, 18, 21, and 24 h post-inoculation. Testing the total virus yield infectivity through virus titration and antigenicity through estimation of complement fixing titer and 146S content and evaluation of the potency of the vaccine prepared from the different cultivation systems were done.

Results: The results showed that the FMDV titer of serotype A, O, and SAT-2 obtained from the roller cultivation system showed the highest level followed by suspension cultivation system then the rawx cultivation system. The FMDV titer showed its highest level at 21 h post-inoculation in all the cultivation systems and then decline at 24 h post-inoculation. The antigenicity reached its highest value content at 18 h post-inoculation either by complement fixation test or by quantifying the 146S intact virion. Montanide ISA 206 oil inactivated trivalent vaccines were prepared from the tested serotypes (A Iran O5. O Panasia and SAT-2/EGY/2012) harvested at 18 h post-inoculation from the 3 culture systems. The results of tracing the antibody response showed that the mean antibody response from the roller cultivation system start its protective antibody titer earlier at 2 weeks post-vaccination (WPV) than the vaccine prepared from the other two cultivation system and the immune protection period lasts longer for 36 WPV for the roller cultivation system vaccine than the other two cultivation systems.

Conclusion: The best cultivation system used for the production of FMD vaccine regarding its highest infectivity and antigenicity is the roller system.

Keywords: baby hamster kidney-21 cell culture, foot and mouth disease virus, monolayer tissue culture cells, suspension tissue culture cells.

Analysis of viral protein-2 encoding gene of avian encephalomyelitis virus from field specimens in Central Java region, Indonesia

Research (Published online: 12-01-2016)

5. Analysis of viral protein-2 encoding gene of avian encephalomyelitis virus from field specimens in Central Java region, Indonesia - Aris Haryanto, Ratna Ermawati, Vera Wati, Sri Handayani Irianingsih and Nastiti Wijayanti

Veterinary World, 9(1): 25-31

   doi: 10.14202/vetworld.2016.25-31

Abstract

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Aim: Avian encephalomyelitis (AE) is a viral disease which can infect various types of poultry, especially chicken. In Indonesia, the incidence of AE infection in chicken has been reported since 2009, the AE incidence tends to increase from year to year. The objective of this study was to analyze viral protein 2 (VP-2) encoding gene of AE virus (AEV) from various species of birds in field specimen by reverse transcription polymerase chain reaction (RT-PCR) amplification using specific nucleotides primer for confirmation of AE diagnosis.

Materials and Methods: A total of 13 AEV samples are isolated from various species of poultry which are serologically diagnosed infected by AEV from some areas in central Java, Indonesia. Research stage consists of virus samples collection from field specimens, extraction of AEV RNA, amplification of VP-2 protein encoding gene by RT-PCR, separation of RT-PCR product by agarose gel electrophoresis, DNA sequencing and data analysis.

Results: Amplification products of the VP-2 encoding gene of AEV by RT-PCR methods of various types of poultry from field specimens showed a positive results on sample code 499/4/12 which generated DNA fragment in the size of 619 bp. Sensitivity test of RT-PCR amplification showed that the minimum concentration of RNA template is 127.75 ng/μl. The multiple alignments of DNA sequencing product indicated that positive sample with code 499/4/12 has 92% nucleotide homology compared with AEV with accession number AV1775/07 and 85% nucleotide homology with accession number ZCHP2/0912695 from Genbank database. Analysis of VP-2 gene sequence showed that it found 46 nucleotides difference between isolate 499/4/12 compared with accession number AV1775/07 and 93 nucleotides different with accession number ZCHP2/0912695.

Conclusions: Analyses of the VP-2 encoding gene of AEV with RT-PCR method from 13 samples from field specimen generated the DNA fragment in the size of 619 bp from one sample with sample code 499/4/12. The sensitivity rate of RT-PCR is to amplify the VP-2 gene of AEV until 127.75 ng/μl of RNA template. Compared to Genbank databases, isolate 499/4/12 has 85% and 92% nucleotide homology.

Keywords: avian encephalomyelitis, reverse transcription polymerase chain reaction, viral protein 2 gene.