Immunopathological response of leukocytes against microfilariae and adult worms in white rats infected with Setaria cervi

Research (Published online: 30-05-2017)

16. Immunopathological response of leukocytes against microfilariae and adult 
worms in white rats infected with Setaria cervi

Sharba Kausar and Wajihullah Khan

Veterinary World, 10(5): 562-568

ABSTRACT

Aim: Aim of this study was to see the immunopathological changes against the microfilariae (Mf) and adult worms of a bovine filarid, Setaria cervi in the tissues of vital organs of experimentally infected white rats. The effect of diethylcarbamazine (DEC) was also observed on the Mf, as leukocytes especially lymphocytes produce immunoglobulins which opsonize and increase the efficacy of DEC against circulating Mf. Effect of this drug was also assessed on liver enzymes in the microfilaremic rats.

Materials and Methods: Microfilaremia was established by implanting adult worms intraperitoneally and by the infusion of Mf recovered from the uterus of gravid female worms. DEC was administered orally for six consecutive days in the rats having patent infection. Differential leukocyte count was recorded every 3rd day, and liver enzymes were estimated every 10th day in both treated and untreated rats. Pathological changes were observed in HE stained sections of vital organs where Mf or adult worms were trapped.

Results: Destruction and reduction in microfilarial density were noticed in microfilaremic rats treated with DEC. Trapped Mf and embedded worms revealed heavy cellular infiltrations by defensive cells which surrounded and attached with the body surface of the Mf as well as adult worms for their destruction and piece meal clearance. Immune-mediated pathology was observed in the tissue sections of lungs, spleen, and liver. Liver enzymes were elevated during the period of higher parasitemia.

Conclusion: There was a moderate level of immunopathology against the Mf and adult worms by the leukocytes in experimentally infected microfilaremic rats. Mf were in the process of degeneration where they got trapped. Moderate increase in liver enzyme was noticed which was slightly more in untreated group. Although a fraction of Mf gets killed in the peritoneum, majority of them successfully enter the systemic circulation and survive for about 54 days, which is sufficient enough for conducting immunological and chemotherapeutic studies.

Keywords: differential leukocyte count, liver enzymes, microfilariae, pathology, Setaria cervi, white rats.

Prevalence of multiple drug resistant Streptococcus suis in and around Guwahati, India

Research (Published online: 28-05-2017)

15. Prevalence of multiple drug resistant Streptococcus suis in and around Guwahati, India

Mrinalee Devi, Jyoti B. Dutta, Swaraj Rajkhowa, Dhireswar Kalita, Girindra Kumar Saikia, Bipin Chandra Das,

Razibuddin Ahmed Hazarika and Gauranga Mahato

Veterinary World, 10(5): 556-561

ABSTRACT

Aim: This study was conducted to determine the prevalence and antimicrobial susceptibility of Streptococcus suis and their resistance patterns isolated from both clinically healthy carriers and diseased pigs in and around Guwahati, Assam, India.

Materials and Methods: A total of 497 samples were collected during October, 2012, to April, 2014, from clinically healthy (n=67) and diseased (n=230) pigs of varying age and either sex maintained under organized and unorganized farming systems. Samples were processed for isolation and identification of S. suis by biochemical characterization and polymerase chain reaction targeting the housekeeping gene glutamate dehydrogenase. In vitro antimicrobial susceptibility of the recovered isolates against nine antibiotic groups comprising 17 antimicrobial agents was studied by standard method.

Results: Of the 497 samples examined, 7 (1.41%) isolates were confirmed to be S. suis of which 5 (1.87%) and 2 (0.87%) were derived from clinically healthy and diseased pigs, respectively. All the isolates were susceptible to gentamicin, amikacin, and erythromycin (100%) followed by the penicillin group and enrofloxacin (85.71%), ceftriaxone, doxycycline HCL, ofloxacin and chloramphenicol (71.43%), to kanamycin, clindamycin and co-trimoxazole (42.85%). The isolates showed least susceptibility to cefalexin, tetracycline and streptomycin (28.57%). All the five S. suis isolates from clinically healthy pigs were susceptible to penicillin G, amoxyclav, doxycycline HCl, gentamicin, amikacin and erythromycin, 80.00% isolates susceptible to ampicillin, enrofloxacin and ofloxacin, 60.00% to ceftriaxone, kanamycin and chloramphenicol, 40% to cefalexin, tetracycline, clindamycin and co-trimoxazole, respectively. Only 20.00% isolates were susceptible to streptomycin. Both the isolates recovered from diseased pigs were susceptible to ampicillin, ceftriaxone, gentamicin, amikacin, enrofloxacin, erythromycin, and clindamycin. On the other hand, both the isolates were resistant to cefalexin, tetracycline, doxycycline HCL, and kanamycin. Altogether five different resistance patterns (multi-drug resistance) were observed. Of the seven S. suis isolates, two isolates were susceptible to all the 17 antimicrobial agents, one isolate was resistant to four antimicrobial agents, two isolates to seven agents, one isolate to nine agents, and one isolate exhibited resistance to 14 antimicrobial agents.

Conclusion: This study was conducted to determine the prevalence of S. suis in clinically healthy and diseased pigs and their antimicrobial susceptibility patterns. All the isolates were susceptible to gentamicin, amikacin and erythromycin, and most of them were resistant to cefalexin, tetracycline and streptomycin. Five different patterns of antimicrobial resistance (multi-drug resistance) were observed.

Keywords: antimicrobial resistance, emerging zoonotic pathogen, multiple drug resistance, pathogen prevalence, Streptococcus suis.

Efficacy and efficiency of poultry carcass composting using different mechanical mixing equipment for avian influenza outbreaks

Research (Published online: 27-05-2017)

4. Efficacy and efficiency of poultry carcass composting using different mechanical mixing equipment for avian influenza outbreaks - Jennifer Elizabeth Keaten and Mark Hutchinson

International Journal of One Health, 3: 19-27

  doi: 10.14202/IJOH.2017.19-27

Abstract

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Background and Aim: Avian influenza (AI) is a viral disease that caused the largest animal disease outbreak in the history of US agriculture. There are several disposal methods of AI infected poultry carcasses available in the US, which include on-site burial, landfill, incineration, rendering, and composting. Of these methods, composting is the most environmentally friendly and poses a low risk for biosecurity. The United States Department of Agriculture (USDA) has developed a comprehensive plan for composting AI infected carcasses. The current protocols have the potential for areas of anaerobic pockets within the windrow due to inadequate mixing and the large carcass size of whole birds. This could lead to ineffective virus neutralization or prolonged composting times and higher resource costs. The purpose of this project was to determine if using a horizontal mixer (HM) wagon to mix composting ingredients or a vertical mixer (VM) wagon to mix and cut up the compositing ingredients is an economical and timely means to accelerate the tissue break-down and obtain optimal temperatures for poultry carcass composting during an AI outbreak.

Materials and Methods: A replicated trial with three treatments, HM, conventional layering (CL) and VM, and three replications was initiated at the Compost Research and Education Center part of the University of Maine Forest and Agricultural Experimental Station called High Moor Farm. Daily temperatures and screened core sample weights (screen weights) on day 0, 16, and 30 were recorded for each of the compost piles. The time to build each replication was recorded and used to help calculate the cost of each method. Data on equipment, carbon material and labor costs were collected from private contractors from the 2014 to 2016 highly pathogenic AI (HPAI) outbreak and used to compare costs between methods.

Results: All treatment methods reached USDA protocol temperatures to neutralize the HPAI virus. Screen weights for both the VM and HM treatments were lower than the CL treatment. Screen weights decreased significantly from day 0 to day 16 for the VM and HM treatments with no significant change from day 16 to day 30. When comparing costs, the mixer wagon methods were the more cost effective than the CL method when using high volume equipment.

Conclusion: The data from this study support the use of a mixer wagon to reduce particle size and mix ingredients for more timely and effective composting of poultry carcasses.

Keywords: carcass management, compost, high pathogenic avian influenza, poultry.

Single-nucleotide polymorphism-based genetic diversity analysis of the Kilakarsal and Vembur sheep breeds

Research (Published online: 26-05-2017)

14. Single-nucleotide polymorphism-based genetic diversity analysis of the Kilakarsal and

Vembur sheep breeds

Rathinasamy Selvam, Nagarajan Murali, A. Kannan Thiruvenkadan, Ramesh Saravanakumar,

Gurusamy Ponnudurai and Thilak Pon Jawahar

Veterinary World, 10(5): 549-555

ABSTRACT

Aim: The present study was thus undertaken to analyze the genetic diversity of Kilakarsal and Vembur sheep breeds using single-nucleotide polymorphism (SNP) markers within Toll-like receptor (TLR) 3, 5, 6, 9, and 10 genes.

Materials and Methods: Competitive allele-specific polymerase chain reaction (PCR)-based end-point genotyping was performed using real-time PCR to type the SNPs. Allele discrimination module implemented in real-time PCR was utilized to call the genotypes based on fluorescence intensity recorded for each of the two alleles. Basic diversity indices, namely, gene frequencies, observed heterozygosity, expected heterozygosity, and inbreeding coefficient (FIS), and testing for Hardy- Weinberg equilibrium (HWE) were estimated using package for elementary analysis of SNP data software program.

Results: Of the 25 SNPs, 22 were found to be polymorphic, whereas two SNPs, namely, TLR3_1081_AC and TLR9_2036_CT, were monomorphic in both Kilakarsal and Vembur sheep populations. The SNP TLR10_1180_AG was monomorphic in Kilakarsal but polymorphic in Vembur sheep. The observed heterozygosities were estimated as 0.289 and 0.309 in Kilakarsal and Vembur sheep, respectively, whereas the expected heterozygosity values were 0.305 and 0.309 in the two breeds, respectively. The overall mean FIS was 0.107 ranging from -0.005 to 0.241 in Kilakarsal sheep and -0.047 ranging from -0.005 to 0.255 in Vembur sheep. In Kilakarsal sheep, the test for HWE revealed TLR9_1308_GC SNP locus with significant deviation (p<0.05) due to heterozygosity deficit. In Vembur sheep, TLR10_82_CT and TLR10_292_CG loci showed significant deviation (p<0.05) due to heterozygosity excess. Other SNP loci did not deviate from HWE (p>0.05) revealing that the population was in HWE proportions.

Conclusion: The SNP markers within five TLR genes (TLR3, TLR5, TLR6, TLR9, and TLR10) utilized for genotyping in this study were highly polymorphic in Kilakarsal and Vembur breeds of sheep. This study on the genetic diversity analysis of the Kilakarsal and Vembur sheep breeds revealed considerable genetic variation within the breeds and it can be utilized to improve desirable traits.

Keywords: allele discrimination module, competitive allele-specific polymerase chain reaction, Kilakarsal, single-nucleotide polymorphism, Toll-like receptor genes, Vembur.

The positive expression of genotype VII Newcastle disease virus (Malaysian isolate) in Japanese quails (Coturnix coturnix japonica)

Research (Published online: 25-05-2017)

13. The positive expression of genotype VII Newcastle disease virus (Malaysian isolate) in

Japanese quails (Coturnix coturnix japonica)

Lizma Felisha Mazlan, Noor Farhana Bachek, Siti Nor Azizah Mahamud, Lokman Hakim Idris, Tan Sheau Wei,

Abdul Rahman Omar and Mohd Hezmee Mohd Noor

Veterinary World, 10(5): 542-548

ABSTRACT

Aim: Genotype VII Newcastle disease virus (NDV) is the most predominant NDV strains that circulating in Malaysia; thus, this study was aimed to determine the susceptibility of Japanese quails toward genotype VII NDV. Clinical signs, gross pathological lesions of organs, positive detection of virus in organs and cloacal swabs, as well as the expression of the antibody titer, were used as parameters to assess the susceptibility of Japanese quails following infection of genotype VII NDV.

Materials and Methods: About 20 quails were divided into three groups (n=8 for Groups A and B; n=4 for the control group). The quails in the Groups A and B were infected via intraocular route with 0.03 ml of 103.5 ELD50 and 107.0 ELD50 of NDV strain IBS 002, respectively, while the control group received 1x phosphate-buffered saline. Cloacal swabs and necropsy were taken on day 7 post-infection for all quails were subjected to one-step reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) for detection of virus and examination for gross pathological lesion, respectively. Blood serums of infected quails were taken on day 10, 14, and 21 post-day infections and were subjected for hemagglutination inhibition (HI) assay.

Results: Depression and ruffled feathers, trachea rales, leg paralysis, and torticollis were shown in some of the quails in both infected groups. Based on statistical analysis, there was no significant difference (p>0.05) in clinical signs between the infected groups. The results for RT-qPCR were found to be negative for all groups, and no gross pathological lesions of organs observed for quails in both infected groups. Trachea, proventriculus, and cecal tonsil were taken for the detection of NDV by RT-qPCR, and some of the organ samples showed positive detection of virus in both infected groups. HI assay showed an increase in mean titers of antibody across time and between infected groups.

Conclusion: In summary, Japanese quails are susceptible to genotype VII NDV based on parameters assessed.

Keywords: infections, intraocular, Japanese quails, Newcastle disease virus.

Standardization of dot-enzyme-linked immmunosorbent assay for the diagnosis of bovine visceral schistosomiasis

Research (Published online: 21-05-2017)

12. Standardization of dot-enzyme-linked immmunosorbent assay for the diagnosis of bovine visceral schistosomiasis

Kommu Sudhakar, G. S. Sreenivasa Murthy and Gaddam Rajeshwari

Veterinary World, 10(5): 536-541

ABSTRACT

Aim: Bovine visceral schistosomiasis has been reported as an important disease entity as it affects animal health, productivity, causes economic losses due to liver condemnation, and produces a high morbidity. This study was conducted to standardize an easy, reliable dot-enzyme-linked immmunosorbent assay (ELISA) for the diagnosis of visceral schistosomiasis caused by Schistosoma spindale and to know the prevalence rate in and around Hyderabad.

Materials and Methods: A dot-ELISA was standardized in the laboratory using whole worm antigen (WWA) and excretory-secretory antigen (ESA) of S. spindale. The standardized test was used for the diagnosis of bovine visceral schistosomiasis at field level. The sensitivity and specificity of the test was compared with counter current immunoelectrophoresis. In total, 288 sera (125 cattle and 163 buffalo) were screened by dot-ELISA.

Results: The dot-ELISA detected 32.63% of infection (94/288) using WWA and 40.62% of infection (117/288) using ESA. In cattle, the prevalence rate was 32.80% (41/125) using WWA and 40.80% (51/125) of infection. Similarly, in buffaloes, the prevalence rate was 32.51% (53/163) using WWA and 40.49% (66/163) of infection using ESA. The overall sensitivity of dot-ELISA was 76.74% and 80.48% with WWA and ESA, respectively, and specificity was 73.3% and 78.57% in WWA and ESA, respectively.

Conclusion: As ante-mortem diagnosis of visceral schistosomiasis is difficult in subclinical conditions, dot-ELISA can be used as a reliable immunodiagnostic test for diagnosis at field level.

Keywords: dot-enzyme-linked immmunosorbent assay, excretory-secretory antigen, Schistosoma spindale, whole worm antigen.

Development of indirect enzyme-linked immunosorbent assay for diagnosis of canine leptospirosis

Research (Published online: 20-05-2017)

11. Development of indirect enzyme-linked immunosorbent assay for diagnosis of canine leptospirosis

A. Sathiyamoorthy, G. Selvaraju, K. M. Palanivel and P. Srinivasan

Veterinary World, 10(5): 530-535

ABSTRACT

Aim: This study was taken up to develop an indirect enzyme-linked immunosorbent assay (i-ELISA) for screening antibodies against Leptospira spp. in canines.

Materials and Methods: An i-ELISA was developed using outer membrane protein extracted from Leptospira interrogans serovar canicola used for coating the well with concentration of 0.5 μg/μl. A total of 250 serum samples from clinically affected and apparently healthy dogs were collected along with relevant epidemiological data at Teaching Veterinary Clinical Complex, Veterinary College and Research Institute, Namakkal, and subjected to i-ELISA.

Results: Out of 250 sera samples, 140 (56.00%) were found to be positive by i-ELISA. All the sera samples were subjected to microagglutination test (MAT) with panel of 12 different serovars. A total of 71 (28.40%) sera samples were positivity to MAT excluding the sera samples positive to L. interrogans serovars canicola and icterohaemorrhagiae in vaccinated dogs. Sensitivity and specificity of i-ELISA were higher in compared with MAT was 91.54% and 58.10%, respectively.

Conclusion: An indirect ELISA developed for the detection of canine antileptospiral antibodies proved to be highly sensitive, rapid and easy to perform and overcome the drawbacks of MAT.

Keywords: canine leptospirosis, indirect enzyme-linked immunosorbent assay, outer membrane protein, Leptospira canicola, Triton X-114 extraction.