Cloning and sequence analysis of a partial CDS of leptospiral ligA gene in pET-32a - Escherichia coliDH5α system

Research (Published online: 30-04-2018)

23. Cloning and sequence analysis of a partial CDS of leptospiral ligA gene in pET-32a - Escherichia coliDH5α system

Manju Soman, Mangattuparambil Mini, Siju Joseph, Jobin Thomas, Nirmal Chacko, T. G. Sumithra, R. Ambily, Binu K. Mani and Rinsha Balan

Veterinary World, 11(4): 557-561

ABSTRACT

Aim: This study aims at cloning, sequencing, and phylogenetic analysis of a partial CDS of ligA gene in pET-32a - Escherichia coli DH5α system, with the objective of identifying the conserved nature of the ligA gene in the genus Leptospira.

Materials and Methods: A partial CDS (nucleotide 1873 to nucleotide 3363) of the ligA gene was amplified from genomic DNA of Leptospira interrogans serovar Canicola by polymerase chain reaction (PCR). The PCR-amplified DNA was cloned into pET-32a vector and transformed into competent E. coli DH5α bacterial cells. The partial ligA gene insert was sequenced and the nucleotide sequences obtained were aligned with the published ligA gene sequences of other Leptospira serovars, using nucleotide BLAST, NCBI. Phylogenetic analysis of the gene sequence was done by maximum likelihood method using Mega 6.06 software.

Results: The PCR could amplify the 1491 nucleotide sequence spanning from nucleotide 1873 to nucleotide 3363 of the ligA gene and the partial ligA gene could be successfully cloned in E. coli DH5α cells. The nucleotide sequence when analyzed for homology with the reported gene sequences of other Leptospira serovars was found to have 100% homology to the 1910 bp to 3320 bp sequence of ligA gene of L. interrogansstrain Kito serogroup Canicola. The predicted protein consisted of 470 aminoacids. Phylogenetic analysis revealed that the ligA gene was conserved in L. interrogans species.

Conclusion: The partial ligA gene could be successfully cloned and sequenced from E. coli DH5α cells. The sequence showed 100% homology to the published ligA gene sequences. The phylogenetic analysis revealed the conserved nature of the ligA gene. Further studies on the expression and immunogenicity of the partial LigA protein need to be carried out to determine its competence as a subunit vaccine candidate.

Keywords: cloning, Escherichia coli DH5α, Leptospira, ligA, pET-32a, phylogenetic tree.

Prevalence and diversity of gastrointestinal helminths in free-ranging Asian house shrew (Suncus murinus) in Bangladesh

Research (Published online: 30-04-2018)

22. Prevalence and diversity of gastrointestinal helminths in free-ranging Asian house shrew (Suncus murinus) in Bangladesh

Mizanur Rahman, Shariful Islam, Md. Masuduzzaman, Mahabub Alam, Mohammad Nizam Uddin Chawdhury, Jinnat Ferdous, Md. Nurul Islam, Mohammad Mahmudul Hassan, Mohammad Alamgir Hossain and Ariful Islam

Veterinary World, 11(4): 549-556

ABSTRACT

Background and Aim: Asian house shrew (Suncus murinus), a widely distributed small mammal in the South Asian region, can carry helminths of zoonotic importance. The aim of the study was to know the prevalence and diversity of gastrointestinal (GI) helminths in free-ranging Asian house shrew (S. murinus) in Bangladesh.

Materials and Methods: A total of 86 Asian house shrews were captured from forest areas and other habitats of Bangladesh in 2015. Gross examination of the whole GI tract was performed for gross helminth detection, and coproscopy was done for identification of specific eggs or larvae.

Results: The overall prevalence of GI helminth was 77.9% (67/86), with six species including nematodes (3), cestodes (2), and trematodes (1). Of the detected helminths, the dominant parasitic group was from the genus Hymenolepis spp. (59%), followed by Strongyloides spp. (17%), Capillaria spp. (10%), Physaloptera spp. (3%), and Echinostoma spp. (3%).

Conclusion: The finding shows that the presence of potential zoonotic parasites (Hymenolepis spp. and Capillaria spp.) in Asian house shrew is ubiquitous in all types of habitat (forest land, cropland and dwelling) in Bangladesh. Therefore, further investigation is crucial to examine their role in the transmission of human helminthiasis.

Keywords: Asian house shrew, Bangladesh, gastrointestinal helminths, prevalence, Suncus murinus.

Effect of Doublesynch and Estradoublesynch protocols on estrus induction, conception rate, plasma progesterone, protein, and cholesterol profile in anestrus Gir heifers

Research (Published online: 27-04-2018)

21. Effect of Doublesynch and Estradoublesynch protocols on estrus induction, conception rate, plasma progesterone, protein, and cholesterol profile in anestrus Gir heifers

N. J. Chaudhary, D. M. Patel, A. J. Dhami, K. B. Vala, K. K. Hadiya and J. A. Patel

Veterinary World, 11(4): 542-548

ABSTRACT

Aim: This study aimed to evaluate the efficacy of Doublesynch and Estradoublesynch protocols on estrus induction, conception rates, plasma progesterone, protein, and cholesterol profile in anestrus Gir heifers.

Materials and Methods: In this study, 50 pubertal anestrus Gir heifers were selected from the field and farm conditions. The heifers were dewormed (injection ivermectin, 100 mg, s/c) and supplemented with minerals and vitamins (injection organic phosphorus 800 mg and injection Vitamin AD3E and Biotin 10 ml i/m) and multi-mineral bolus at 1 bolus daily for 7 days. The heifers were randomly divided into three groups: Doublesynch (n=20), Estradoublesynch (n=20), and control (n=10). The animals were monitored for estrus response, estrus interval, behavioral signs, and conception rates after induced/first, second, and third cycle post-treatment. Blood samples were obtained on day 0, day 9, day 12, and on day 12 post-artificial insemination (AI) for determination of plasma progesterone, protein, and cholesterol profile.

Results: The estrus response rate between Doublesynch and Estradoublesynch protocols was similar between treated heifers (85% and 95%). The interval from the second prostaglandin F2α (PGF2α) injection to estrus induction did not differ between the groups (63.87±4.19 vs. 58.27±3.83 h). The conception rates following induced estrus (20% vs. 30%), at the second cycle (23.07% vs. 16.66%), at the third cycle (22.22% vs. 30.00%), and the overall conception rate (45% and 55%) within 27.89±5.75 and 26.45±5.48 days were the same across the treatment groups. The mean plasma progesterone concentrations were significantly (p<0.01) higher on day 9 (second PGF2α injection) and day 12 post-AI compared to day 0 (first PGF2α injection) and the day of fixed-timed artificial insemination. The concentrations were also significantly (p<0.05) higher in conceived than non-conceived heifers on day 9 of treatment and day 12 post-AI in both the protocols. The mean plasma cholesterol concentrations were significantly higher during peak follicular and luteal phases compared to the initial anestrus phase in both the protocols. The values were also higher in non-conceived than conceived animals in both the protocols. The plasma protein profile was not influenced by the sampling days or conceived and non-conceived status.

Conclusion: The results showed that both Doublesynch and Estradoublesynch protocols resulted in similar estrus induction and conception rates with modulation of plasma progesterone and cholesterol profile in anestrus Gir heifers.

Keywords: cholesterol, conception rate, estrus synchronization, Gir heifers, progesterone, proteins, pubertal anestrus.

Copy number variation in livestock: A mini review

Review (Published online: 26-04-2018)

20. Copy number variation in livestock: A mini review

V. Bhanuprakash, Supriya Chhotaray, D. R. Pruthviraj, Chandrakanta Rawat, A. Karthikeyan and Manjit Panigrahi

Veterinary World, 11(4): 535-541

ABSTRACT

Copy number variation (CNV) is a phenomenon in which sections of the genome, ranging from one kilo base pair (Kb) to several million base pairs (Mb), are repeated and the number of repeats vary between the individuals in a population. It is an important source of genetic variation in an individual which is now being utilized rather than single nucleotide polymorphisms (SNPs), as it covers the more genomic region. CNVs alter the gene expression and change the phenotype of an individual due to deletion and duplication of genes in the copy number variation regions (CNVRs). Earlier, researchers extensively utilized SNPs as the main source of genetic variation. But now, the focus is on identification of CNVs associated with complex traits. With the recent advances and reduction in the cost of sequencing, arrays are developed for genotyping which cover the maximum number of SNPs at a time that can be used for detection of CNVRs and underlying quantitative trait loci (QTL) for the complex traits to accelerate genetic improvement. CNV studies are also being carried out to understand the evolutionary mechanism in the domestication of livestock and their adaptation to the different environmental conditions. The main aim of the study is to review the available data on CNV and its role in genetic variation among the livestock.

Keywords: copy number variation, copy number variation regions, livestock, single nucleotide polymorphisms, quantitative trait loci.

Molecular detection of Leptospira spp. from canine kidney tissues and its association with renal lesions

Research (Published online: 25-04-2018)

19. Molecular detection of Leptospira spp. from canine kidney tissues and its association with renal lesions

Biswajit R. Dash, Vitthal S. Dhaygude, Prashant D. Gadhave, Kaustubh V. Garud and Dattatarya P. Kadam

Veterinary World, 11(4): 530-534

ABSTRACT

Aim: The study aimed to detect the prevalence of Leptospira spp. in kidney tissues collected during necropsy and to establish its association with renal lesions in dogs of Mumbai region.

Materials and Methods: Kidney tissues from 40 dogs were collected during necropsy after gross examination and then fixed in neutral buffered formalin and Bouin's fluid for histopathology and histochemistry, respectively. Kidney tissues were also collected for the detection of Leptospira spp. by polymerase chain reaction (PCR) in a sterile container and stored at -80°C until further processing.

Results: Of 40 cases studied, 13 (32.5%) cases showed lesions of nephritis of varying histotype and severity. Glomerulonephritis was reported as the most common type of nephritis in 9 (69.23%) cases, and interstitial nephritis was recorded in 4 (30.76%) cases. Chronic and acute interstitial nephritis was observed in two cases each. Renal failure as a cause of death was found in 7 (17.5%) dogs. Of a total of 40 cases, 9 were found positive for pathogenic Leptospira spp. genome by PCR. However, of nine PCR-positive cases, only four cases showed lesions in kidneys as glomerulonephritis and interstitial nephritis in two cases each. The rest five cases positive for Leptospira spp. by PCR did not show any appreciable lesions in the kidneys.

Conclusion: Leptospiral DNA was detected in 9 (22.5%) cases by PCR. Of these nine cases, only four cases showed renal lesions. Other five cases which were positive for Leptospira spp. by PCR did not show any appreciable gross and microscopic lesions in the kidneys which might be carriers for Leptospira spp. Considering variable reports on types of nephritis in Leptospira spp. infection and also the prevalence of non-pathogenic Leptospira spp., it is important to conduct an extensive study on the prevalence of Leptospira spp. and its association with renal lesions involving batteries of tests.

Keywords: histopathology, kidneys, Leptospira spp., nephritis.

Pseudopregnancy in goats: Sonographic prevalence and associated risk factors in Khartoum State, Sudan

Research (Published online: 23-04-2018)

18. Pseudopregnancy in goats: Sonographic prevalence and associated risk factors in Khartoum State, Sudan

Areeg Mohamed Almubarak, Naglaa Abd Elhakeem Abass, Majdi Elnaim Badawi, Mohamed Tagelddin Ibrahim, Abdelhamid Ahmed Elfadil and Rihab Mohamed Abdelghafar

Veterinary World, 11(4): 525-529

ABSTRACT

Aim: This study was conducted to estimate the prevalence of pseudopregnancy in goats and to investigate potential risk factors associated with the condition in Khartoum State.

Materials and Methods: A cross-sectional study was carried out from March 2015 to February 2016. A total of 378 female goats which presented to the Veterinary Teaching Hospital, College of Veterinary Medicine, Sudan University of Science and Technology, for routine ultrasonographic pregnancy diagnosis were examined. Ultrasound scanning was performed using a real-time scanner equipped with dual-frequency (3.5-5 MHz) curvilinear transducer.

Results: The results showed that the prevalence of pseudopregnancy in goats in Khartoum State was 10.6%. Risk factors such as general body condition (χ2=5.974; p=0.05), age (χ2=11.760; p=0.0129), type of estrus (χ2=12.794; p=0.000), and previous reproductive performance (χ2=13.397; p=0.020) showed significant association (p≤0.05) with the occurrence of pseudopregnancy in the univariate analysis. Breed (χ2=12.627; p=0.082), milk yield (χ2=5.951; p=0.114), type of feeding (χ2=1.721; p=0.190), season (χ2=2.661; p=0.264), locality (χ2=7.66; p=0.264), parity number (χ2=0.451; p=0.767), and rearing system (χ2=1.593; p=0.451) were not significantly associated with pseudopregnancy.

Conclusion: The prevalence of pseudopregnancy in goats in Khartoum State was 10.6%. Pseudopregnancy in goats is significantly associated with age, type of estrus, general body condition, and previous reproductive performance. This study showed for the first time that pseudopregnancy is a real reproductive problem in goats in Khartoum State.

Keywords: goat, hydrometra, prevalence, risk factors, Sudan, ultrasound.

Isolation, identification, and serotyping of Avibacterium paragallinarum from quails in Indonesia with typical infectious coryza disease symptoms

Research (Published online: 23-04-2018)

17. Isolation, identification, and serotyping of Avibacterium paragallinarum from quails in Indonesia with typical infectious coryza disease symptoms

Agnesia Endang Tri Hastuti Wahyuni, Charles Rangga Tabbu, Sidna Artanto, Dwi Cahyo Budi Setiawan and Sadung Itha Rajaguguk

Veterinary World, 11(4): 519-524

ABSTRACT

Background and Aim: Infectious coryza (IC) or snot is an infectious upper respiratory disease affecting chickens and birds, including quails, and it is caused by Avibacterium paragallinarum. The symptoms of IC are facial swelling, malodorous nasal discharge, and lacrimation. This study aimed to isolate, identify, and serotype the A. paragallinarum of snot in quails and to determine the sensitivity and resistance to several antibiotics.

Materials and Methods: Nine quails from Yogyakarta, Indonesia with typical snot disease symptoms were used in this study. The nasal swab was obtained and directly streaked onto a chocolate agar plate and blood agar plate (BAP), then incubated in 5% CO2 at 37°C for 24-48 h. Staphylococcus spp. was cross-streaked onto the BAP to show the satellite growth. The observation of the morphology of the suspected colony, Gram staining, and biochemical tests (catalase test, oxidase test, urease test, peptone test, and carbohydrate fermentation such as maltose, mannitol, lactose, and sorbitol) are done to identify the species of bacteria. This research also detects the serovar of A. paragallinarum using hemagglutination inhibition test. The antibiotic sensitivity tests were also performed using several antibiotics against five A. paragallinarum isolates that were cultured on Mueller-Hinton Agar and added with antibiotic discs, then incubated in 5% CO2 at 37°C for 24-48 h.

Results: Five isolates out of nine suspected isolates (55.5%) were A. paragallinarum. The growth of isolates from quails did not depend on the nicotinamide adenine dinucleotide (NAD) (NAD-independent). Sensitivity test was done using the five identified A. paragallinarum isolates, results showed that they were 100% sensitive to amoxicillin (AMC) and ampicillin (AMP); 100% resistant toward amikacin (AK), erythromycin (E), gentamycin (CN), and tetracycline (TE); 80% resistant toward kanamycin (K) and trimethoprim (W); 60% resistant toward chloramphenicol (C); and 20% toward enrofloxacin (ENR). The antibiotics that have an intermediate sensitivity (in between sensitive and resistant) were ENR and K, 80% and 20%, respectively. Three out of five A. paragallinarum isolates were identified as serovar B of A. paragallinarum using HI test.

Conclusion: Five out of nine isolates (55.5%) from quails with typical IC disease symptoms identified as A. paragallinarum and sensitive toward AMC and AMP. Three out of five A. paragallinarum isolates were identified as serovar B.

Keywords: antibiotic sensitivity test, Avibacterium paragallinarum, infectious coryza, nicotinamide adenine dinucleotide-independent.

Seroprevalence and risk factors of caprine brucellosis in Khartoum state, Sudan

Research (Published online: 19-04-2018)

16. Seroprevalence and risk factors of caprine brucellosis in Khartoum state, Sudan

Eman Mohamed-Ahmed Mohamed, Abdelhamid Ahemd Mohamed Elfadil, Enaam Mohamed El-Sanousi, Hatim Hamad Ibrahaem, Saad El-Tiab Mohamed-Noor, Mohamed Abdelsalam Abdalla and Yassir Adam Shuaib

Veterinary World, 11(4): 511-518

ABSTRACT

Aim: This cross-sectional study was conducted from April to July 2012 in Khartoum state, Sudan, to determine the seroprevalence of brucellosis in goats and to investigate potential risk factors associated with this disease.

Materials and Methods: A total of 307 serum samples were collected from both sexes of goats in four different localities and were subjected to testing for brucellosis using rose bengal plate test (RBPT), serum agglutination test (SAT), and competitive enzyme-linked immunosorbent assay (cELISA).

Results: The overall seroprevalence was 11.4% (n=35) with a 95% confidence interval (CI) ranging from 7.80 to 15.0. Out of these 35 RBPT-positive samples, the positivity of 18 and 17 were confirmed by SAT and cELISA, respectively. A significant statistical variation was observed between brucellosis seroprevalences in goats purchased from local animal markets and goats that were raised at the farm. Conversely, such significant variations were not observed among the categories of other risk factors with seroprevalences ranging from 3.0% (95% CI between 0.40 and 7.20) to 16.3% (95% CI between 10.4 and 22.3). Location (χ2=9.33, df=3, p=0.02), breed (χ2=3.52, df=1, p=0.05), herd size (χ2=6.59, df=2, p=0.03), and herd expansion (χ2=5.39, df=1, p=0.02) were associated with RBPT-positive status for brucella in the two-tailed Chi-square test. In addition, Sharq an-Nil locality and goats raised at the farm had increased odds of being RBPT positive.

Conclusion: Brucellosis was detected in goats in all surveyed localities. An effort should be made to educate goat owners/herders about brucellosis as well as about the importance of vaccination.

Keywords: brucellosis, goats, risk factors, rose bengal plate test, seroprevalence, Sudan.

Viability test of fish scale collagen (Oshpronemus gouramy) on baby hamster kidney fibroblasts-21 fibroblast cell culture

Research (Published online: 18-04-2018)

15. Viability test of fish scale collagen (Oshpronemus gouramy) on baby hamster kidney fibroblasts-21 fibroblast cell culture

Chiquita Prahasanti, Denny Tri Wulandari and Noer Ulfa

Veterinary World, 11(4): 506-510

Aim: This study aims to examine the toxicity of collagen extracted from gouramy fish scales (Oshpronemus gouramy) by evaluating its viability against baby hamster kidney fibroblasts-21.

Materials and Methods: Collagen was extracted from gouramy fish scales (O. gouramy) with 6% acetic acid. Its results were analyzed using Fourier-transform infrared spectroscopy and freeze-dried technique. Its morphology then was analyzed with scanning electron microscope. Afterward, 3-(4.5-dimethylthiazole-2-yl)2.5-diphenyl tetrazolium bromide assay was conducted to compare cells with and without fish scale collagen treatment.

Results: Collagen extracted from gouramy fish scales had no influence statistically on cultured fibroblast cells with a statistical significance (2-tailed) value of 0.754 (p>00025).

Conclusion: Collagen extracted from gouramy fish scales has high viability against BHK21 fibroblast cells.

Keywords: bone graft, collagen, gouramy fish scale, viability, 3-(4.5-dimethylthiazole-2-yl)2.5-diphenyl tetrazolium bromide.

Genetic characterization of 11 microsatellite loci in Egyptian pigeons (Columba livia domestica) and their cross-species amplification in other Columbidae populations

Research (Published online: 17-04-2018)

14. Genetic characterization of 11 microsatellite loci in Egyptian pigeons (Columba livia domestica) and their cross-species amplification in other Columbidae populations

Sherif Ramadan, Ahmed Dawod, Osama El-Garhy, Amira M. Nowier, Marwa Eltanany and Miho Inoue-Murayama

Veterinary World, 11(4): 497-505

ABSTRACT

Aim: This study aimed to analyze the genetic diversity and relationships of 10 Egyptian pigeon populations belonging to Columba livia domestica species using 11 microsatellite markers and to investigate the success of these markers amplification across another eight pigeon species.

Methods: Genomic DNA was isolated from feather samples of 179 pigeon samples from 10 Egyptian breeds: Asfer Weraq (n=14), Austoraly (n=20), Reehani (n=21), Messawed (n=17), Nemssawy (n=27), Otatti (n=12), Morasla (n=17), Tumbler (n=22), Halaby Asfer (n=10), and Karakandy (n=19) in addition to Japanese feral pigeons (n=30). Genotyping was done using 11 specific polymorphic microsatellite makers. Moreover, 37 samples not belonging to C. livia domestica but belonging to another eight pigeon species were genotyped. The polymerase chain reaction (PCR) products were electrophoresed on an ABI 3130xl DNA Sequencer. The basic measures of genetic diversity and phylogenetic trees were computed using bioinformatics software.

Results: Across the 10 studied Egyptian populations, the number of alleles per locus ranged from 3 to 19 and the average number of alleles observed was 9.091. The lowest value of expected heterozygosity (0.373) was obtained for the Reehani breed, and the highest value (0.706) was found for Morasla breed. The overall expected heterozygosity of Egyptian pigeons was 0.548. The FST coefficient which indicates fixation coefficients of subpopulations within the total population for the 11 loci varied from 0.318 to 0.114 with a relatively high mean (0.226). In our study, the FIS showed a relatively high average (0.037). The pairwise Reynolds's genetic distance between the 11 studied pigeon populations recorded lower values between Otatti and Austoraly (0.025) and between Morasla and Japanese feral pigeons (0.054). These results are supported by clustering pattern either by the neighbor-joining phylogenetic tree or by a Bayesian clustering of STRUCTURE with the admixture method.

Conclusion: We confirm the applicability of the CliμD17, CliμT17, CliμD16, CliμD32, CliμT13, CliμD01, PG1, PG2, PG4, PG6, and PG7 microsatellite markers among Egyptian domestic pigeons and across other pigeon species using cross-species amplification method. The information from this study should be useful for genetic characterization and for developing conservation programs of this important species.

Keywords: Egyptian breed, genetic diversity, microsatellite, pigeon.

Radiographic prediction of metallic foreign body penetration in the reticulum of cows and buffaloes

Research (Published online: 16-04-2018)

13. Radiographic prediction of metallic foreign body penetration in the reticulum of cows and buffaloes

Shanib Mehraj Makhdoomi, Vandana Sangwan, and Ashwani Kumar

Veterinary World, 11(4): 488-496

ABSTRACT

Aim: This study aimed to evaluate the role of radiography in the standing (right and left) and recumbent (right) lateral positions for the detection and prediction of metallic foreign body penetration in the reticular wall.

Materials and Methods: A total of 41 bovines (23 cows and 18 buffaloes) having at least one sharp metallic foreign body (>1 cm) detected on reticular radiographs were investigated, and their extent of penetration in the reticular wall was confirmed on the left flank laparorumenotomy.

Results: Of total sharp metallic foreign bodies retrieved on rumenotomy, the maximum percent were detected on the right recumbent radiographic view (75.00% in cows and 57.14% in buffaloes) compared to the right standing (54.38% in cows and 40.42% in buffaloes) and left standing (51.06% in cows and 27.08% in buffaloes) radiographic views. The presence of gas pocket or nodule adjoining a foreign body, faintly visible foreign body, foreign body that appeared partially or completely out of the reticulum, and foreign body that appeared parallel, into, or directed toward the diaphragm indicated a high probability in the prediction of penetrating foreign body in the left standing (100%) followed by the right recumbent (85.71% in cattle and 90% in buffaloes) and right standing (94.74% in cattle and 55.56% in buffaloes) radiographic views.

Conclusion: The right recumbent radiographic view is most reliable to detect sharp metallic foreign bodies in bovine. Buffaloes engulf more number of foreign bodies; however, comparatively, the number of completely or partially penetrating foreign bodies is high in cattle. The hypothesized radiographic parameters for the prediction of penetrability of the metallic foreign body were 100% reliable in the left standing radiographic view in both the species.

Keywords: bovine, cows, cranioventral abdomen, radiograph, reticulum.

Molecular characterization of canine parvovirus variants (CPV-2a, CPV-2b, and CPV-2c) based on the VP2 gene in affected domestic dogs in Ecuador

Research (Published online: 16-04-2018)

12. Molecular characterization of canine parvovirus variants (CPV-2a, CPV-2b, and CPV-2c) based on the VP2 gene in affected domestic dogs in Ecuador

David De la Torre, Eulalia Mafla, Byron Puga, Linda Erazo, Claudete Astolfi-Ferreira and Antonio Piantino Ferreira

Veterinary World, 11(4): 480-487

ABSTRACT

Aim: The objective of this study was to determine the presence of the variants of canine parvovirus (CPV)-2 in the city of Quito, Ecuador, due to the high domestic and street-type canine population, and to identify possible mutations at a genetic level that could be causing structural changes in the virus with a consequent influence on the immune response of the hosts.

Materials and Methods: Thirty-five stool samples from different puppies with characteristic signs of the disease and positives for CPV through immunochromatography kits were collected from different veterinarian clinics of the city. Polymerase chain reaction and DNA sequencing were used to determine the mutations in residue 426 of the VP2 gene, which determines the variants of CPV-2; in addition, four samples were chosen for complete sequencing of the VP2 gene to identify all possible mutations in the circulating strains in this region of the country.

Results: The results revealed the presence of the three variants of CPV-2 with a prevalence of 57.1% (20/35) for CPV-2a, 8.5% (3/35) for CPV-2b, and 34.3% (12/35) for CPV-2c. In addition, complete sequencing of the VP2gene showed amino acid substitutions in residues 87, 101, 139, 219, 297, 300, 305, 322, 324, 375, 386, 426, 440, and 514 of the three Ecuadorian variants when compared with the original CPV-2 sequence.

Conclusion: This study describes the detection of CPV variants in the city of Quito, Ecuador. Variants of CPV-2 (2a, 2b, and 2c) have been reported in South America, and there are cases in Ecuador where CVP-2 is affecting even vaccinated puppies.

Keywords: canine parvovirus, canine parvovirus-2, Ecuador, molecular characterization, variants.

Evaluation of multiplex polymerase chain reaction as an alternative to conventional antibiotic sensitivity test

Research (Published online: 13-04-2018)

11. Evaluation of multiplex polymerase chain reaction as an alternative to conventional antibiotic sensitivity test

K. Rathore, B. Joseph, D. K. Sharma, A. Gaurav, S. K. Sharma, M. Milind, P. Patel, C. Prakash and L. Singh

Veterinary World, 11(4): 474-479

ABSTRACT

Aim: This study was designed to evaluate the potential of the use of multiplex polymerase chain reaction (PCR) as an alternative to conventional antibiotic sensitivity test.

Materials and Methods: Isolates of Staphylococcus aureus (total = 36) from clinical cases presented to Teaching Veterinary Clinical Complex of College of Veterinary and Animal Sciences (CVAS), Navania, Udaipur, were characterized by morphological, cultural, and biochemical methods. Then, the isolates were further subjected to molecular characterization by PCR targeting S. aureus-specific sequence (107 bp). Phenotypic antibiotic sensitivity pattern was analyzed by Kirby Bauer disc diffusion method against 11 commonly used antibiotics in veterinary medicine in and around Udaipur region. The genotypic antibiotic sensitivity pattern was studied against methicillin, aminoglycosides, and tetracycline targeting the gene mecA, aacA-aphD, and tetK by multiplex PCR.

Results: There was 100% correlation between the phenotype and genotype of aminoglycoside resistance, more than 90% correlation for methicillin resistance, and 58.3% in the case tetracycline resistance.

Conclusion: As there is a good correlation between phenotype and genotype of antibiotic resistance, multiplex PCR can be used as an alternative to the conventional antibiotic susceptibility testing, as it can give a rapid and true prediction of antibiotic sensitivity pattern.

Keywords: antimicrobial resistance, genotype, phenotype, Staphylococcus aureus.

Characterization of quinolone-resistant Enterobacteriaceae strains isolated from poultry in Western Algeria: First report of qnrS in an Enterobacter cloacae

Research (Published online: 12-04-2018)

10. Characterization of quinolone-resistant Enterobacteriaceae strains isolated from poultry in Western Algeria: First report of qnrS in an Enterobacter cloacae

Qada Benameur, Hassiba Tali-Maamar, Farida Assaous, Badia Guettou, Meki Boutaiba Benklaouz, Kheira Rahal and Meriem-Hind Ben-Mahdi

Veterinary World, 11(4): 469-473

ABSTRACT

Aim: Multidrug-resistant (MDR) Enterobacteriaceae have frequently been reported, in both human and veterinary medicine, from different parts of the world as a consequence of antibiotic usage. However, there is a lack of published data regarding antimicrobial resistance in non-Escherichia coli (E. coli) Enterobacteriaceae from animals in Algeria. This study aimed to evaluate the frequency of resistance to antibiotics with a focus on quinolones and to investigate the presence of qnr genes in Enterobacteriaceae of poultry origin.

Materials and Methods: A total of 310 samples of poultry origin were collected from 2010 to 2014 from broiler and layer farms and hatcheries located in different geographic areas of Western Algeria (including Mostaganem, Oran, Mascara, Relizane, Chlef, Tiaret, and Tissemsilt). Antimicrobial susceptibility testing was performed using disc diffusion assay. Polymerase chain reaction and sequencing accomplished the characterization of qnr genes (qnrA, qnrB, and qnrS).

Results: A total of 253 Enterobacteriaceae strains were isolated in this study. These isolates exhibited high levels of resistance to quinolones and other families of antibiotics. All the strains isolated in this study were resistant to at least one antibiotic. Among them, 233 (92.09%) were considered MDR. Among the 18 randomly selected nalidixic acid (NA)- resistant Enterobacteriaceae isolates, one E. coli and one Enterobacter cloacaewere carrying qnrS1. By contrast, qnrA and qnrB were not detected in this study.

Conclusion: This is the first report on the identification of the qnrS gene in E. cloacae isolated from animal source in Algeria. Further studies have to be conducted to determine the real prevalence of qnr genes.

Keywords: Algeria, antimicrobial resistance, Enterobacteriaceae, qnrS1.

Prevalence and magnitude of acidosis sequelae to rice-based feeding regimen followed in Tamil Nadu, India

Research (Published online: 11-04-2018)

9. Prevalence and magnitude of acidosis sequelae to rice-based feeding regimen followed in Tamil Nadu, India

Rathinam Murugeswari, Chinnamani Valli, Raman Karunakaran, Venkatasubramanian Leela and Amaresan Serma Saravana Pandian

Veterinary World, 11(4): 464-468

Background and Aim: In Tamil Nadu, a southern state of India, rice is readily available at a low cost, hence, is cooked (cooking akin to human consumption) and fed irrationally to cross-bred dairy cattle with poor productivity. Hence, a study was carried out with the objective to examine the prevalence of acidosis sequelae to rice-based feeding regimen and assess its magnitude.

Materials and Methods: A survey was conducted in all the 32 districts of Tamil Nadu, by randomly selecting two blocks per districts and from each block five villages were randomly selected. From each of the selected village, 10 dairy farmers belonging to the unorganized sector, owning one or two cross-bred dairy cows in early and mid-lactation were randomly selected so that a sample size of 100 farmers per district was maintained. The feeding regimen, milk yield was recorded, and occurrence of acidosis and incidence of laminitis were ascertained by the veterinarian with the confirmative test to determine the impact of feeding cooked rice to cows.

Results: It is observed that 71.5% of farmers in unorganized sector feed cooked rice to their cattle. The incidence of acidosis progressively increased significantly (p<0.05) from 29.00% in cows fed with 0.5 kg of cooked rice to 69.23% in cows fed with more than 2.5 kg of cooked rice. However, the incidence of acidosis remained significantly (p<0.05) as low as 9.9% in cows fed feeding regimen without cooked rice which is suggestive of a correlation between excessive feeding cooked rice and onset of acidosis. Further, the noticeable difference in the incidence of acidosis observed between feeding cooked rice and those fed without rice and limited intake of oil cake indicates that there is a mismatch between energy and protein supply to these cattle. Among cooked rice-based diet, the incidence of laminitis increased progressively (p<0.05) from 9.2% to 37.9% with the increase in the quantum of cooked rice in the diet.

Conclusion: The study points out the importance of protein supplementation in rice-based feeding regimen to set right the mismatched supply between nitrogen and fermentable organic matter in the rumen. This research has practical implications for animal health, welfare, nutrition, and management.

Keywords: acidosis, cooked rice, dairy cows, digestive disturbance, survey.

Effect of Azolla feeding on the growth, feed conversion ratio, blood biochemical attributes and immune competence traits of growing turkeys

Research (Published online: 11-04-2018)

8. Effect of Azolla feeding on the growth, feed conversion ratio, blood biochemical attributes and immune competence traits of growing turkeys

Mayank Shukla, Amitav Bhattacharyya, Pankaj Kumar Shukla, Debashis Roy, Brijesh Yadav and Rajneesh Sirohi

Veterinary World, 11(4): 459-463

Background and Aim: In the recent past, few studies have been carried out in chicken to assess the effect of Azolla meal and raw Azolla feeding on the performance of chicken. If turkeys effectively use unconventional feedstuffs like Azolla without reducing the performance, it will increase the profitability of turkey business. Hence, a study was carried out to evaluate the effect of dried Azolla pinnata vis-a-vis raw Azolla as choice feeding on the growth, feed conversion ratio (FCR), blood biochemical attributes, and immune competence traits of growing turkeys under intensive system.

Materials and Methods: A total of 72, 8-week-old grower turkey poults of black variety were randomly distributed into three dietary treatments having three replicates each with eight birds. The birds of the control group (T1) were fed a basal diet (CP - 19.71% and ME - 2789.79 Kcal/kg), while the other group (T2) and choice-feeding group (T3) were fed 5% of basal diet replaced by dry Azolla powder on DM basis and ad libitum Azolla along with basal diet, respectively.

Results: There was no significant difference among the different groups in the average weekly weight gain during the entire experiment. FCR was significantly better (p<0.05) in the choice-feeding group compared to the other two experimental groups during 8-16 weeks of age. There was no significant difference among the treatment groups in any of the blood biochemical indices except plasma uric acid, which was significantly decreased (p<0.01) in T2 compared to T1 at 16 weeks of age. HA and IgM response to 1% sheep red blood cells (log2 titer) were numerically better in T2 and T3 compared to the T1.

Conclusion: Thus, it may be inferred that choice feeding with Azolla, and basal diet may improve FCR without any adverse effect on blood biochemical attributes and immune competence traits.

Keywords: Azolla, biochemical attributes, body weight, immunity, turkeys.

Standardization and application of real-time polymerase chain reaction for rapid detection of bluetongue virus

Research (Published online: 10-04-2018)

7. Standardization and application of real-time polymerase chain reaction for rapid detection of bluetongue virus

I. Karthika Lakshmi, Kalyani Putty, Satya Samparna Raut, Sunil R. Patil, P. P. Rao, B. Bhagyalakshmi, Y. Krishna Jyothi, B. Susmitha, Y. Vishnuvardhan Reddy, Sowmya Kasulanati, J. Shiva Jyothi and Y. N. Reddy

Veterinary World, 11(4): 452-458

ABSTRACT

Aim: The present study was designed to standardize real-time polymerase chain reaction (PCR) for detecting the bluetongue virus from blood samples of sheep collected during outbreaks of bluetongue disease in the year 2014 in Andhra Pradesh and Telangana states of India.

Materials and Methods: A 10-fold serial dilution of Plasmid PUC59 with bluetongue virus (BTV) NS3 insert was used to plot the standard curve. BHK-21 and KC cells were used for in vitro propagation of virus BTV-9 at a TCID50/ml of 105 ml and RNA was isolated by the Trizol method. Both reverse transcription -PCR and real-time PCR using TaqMan probe were carried out with RNA extracted from virus-spiked culture medium and blood to compare the sensitivity by means of finding out the limit of detection (LoD). The results were verified by inoculating the detected and undetected dilutions onto cell cultures with further cytological (cytopathic effect) and molecular confirmation (by BTV-NS1 group-specific PCR). The standardized technique was then applied to field samples (blood) for detecting BTV.

Results: The slope of the standard curve obtained was -3.23, and the efficiency was 103%. The LoD with RT-PCR was 8.269Ex103 number of copies of plasmid, whereas it was 13 with real-time PCR for plasmid dilutions. Similarly, LoD was determined for virus-spiked culture medium, and blood with both the types of PCR and the values were 103 TCID 50/ml and 104 TCID 50/ml with RT-PCR and 10° TCID 50/ml and 102 TCID 50/ml with real-time PCR, respectively. The standardized technique was applied to blood samples collected from BTV suspected animals; 10 among 20 samples were found positive with Cq values ranging from 27 to 39. The Cq value exhibiting samples were further processed in cell cultures and were confirmed to be BT positive. Likewise, Cq undetected samples on processing in cell cultures turned out to be BTV negative.

Conclusion: Real-time PCR was found to be a very sensitive as well as reliable method to detect BTV present in different types of samples, including blood samples collected from BTV-infected sheep, compared to RT-PCR. The LoD of BTV is likely influenced by sample type, possibly by the interference by the other components present in the sample.

Keywords: bluetongue virus, limit of detection, real-time polymerase chain reaction.

Effects of lipid extraction on nutritive composition of winged bean (Psophocarpus tetragonolobus), rubber seed (Hevea brasiliensis), and tropical almond (Terminalia catappa)

Research (Published online: 10-04-2018)

6. Effects of lipid extraction on nutritive composition of winged bean (Psophocarpus tetragonolobus), rubber seed (Hevea brasiliensis), and tropical almond (Terminalia catappa)

Anuraga Jayanegara, Rakhmad P. Harahap, Richard F. Rozi and Nahrowi

Veterinary World, 11(4): 446-451

ABSTRACT

Aim: This experiment aimed to evaluate the nutritive composition and in vitro rumen fermentability and digestibility of intact and lipid-extracted winged bean, rubber seed, and tropical almond.

Materials and Methods: Soybean, winged bean, rubber seed, and tropical almond were subjected to lipid extraction and chemical composition determination. Lipid extraction was performed through solvent extraction by Soxhlet procedure. Non-extracted and extracted samples of these materials were evaluated for in vitro rumen fermentation and digestibility assay using rumen: Buffer mixture. Parameters measured were gas production kinetics, total volatile fatty acid (VFA) concentration, ammonia, in vitro dry matter (IVDMD) and in vitro organic matter digestibility (IVOMD). Data were analyzed by analysis of variance and Duncan's multiple range test.

Results: Soybean, winged bean, rubber seed, and tropical almond contained high amounts of ether extract, i.e., above 20% DM. Crude protein contents of soybean, winged bean, rubber seed, and tropical almond increased by 17.7, 4.7, 55.2, and 126.5% after lipid extraction, respectively. In vitro gas production of intact winged bean was the highest among other materials at various time point intervals (p<0.05), followed by soybean > rubber seed > tropical almond. Extraction of lipid increased in vitro gas production, total VFA concentration, IVDMD, and IVOMD of soybean, winged bean, rubber seed, and tropical almond (p<0.05). After lipid extraction, all feed materials had similar IVDMD and IVOMD values.

Conclusion: Lipid extraction improved the nutritional quality of winged bean, rubber seed, and tropical almond.

Keywords: Hevea brasiliensis, in vitro rumen, lipid extraction, Psophocarpus tetragonolobus, Terminalia catappa.