Isolation, molecular characterization and prevalence of Clostridium perfringens in sheep and goats of Kashmir Himalayas, India

Research (Published online: 22-12-2017)

14. Isolation, molecular characterization and prevalence of Clostridium perfringens in sheep and goats of Kashmir Himalayas, India

Salik Nazki, Shakil A. Wani, Rafia Parveen, Showkat A. Ahangar, Zahid A. Kashoo, Syed Hamid, Zahoor A. Dar, Tanveer A. Dar and Pervaiz A. Dar

Veterinary World, 10(12): 1501-1507

ABSTRACT

Aim: The study was conducted to report the occurrence of the Clostridium perfringens in sheep and goats of the Kashmir valley for the 1st time and to characterize them molecularly with respect to toxin genes to determine the prevalence of the various toxinotypes.

Materials and Methods: A total of 177 samples (152 from sheep and 25 from goats) collected from healthy, diarrheic animals, and morbid material of animals suspected to have died of enterotoxaemia were screened for C. perfringens toxinotypes. The presumptive positive isolates were confirmed using 16S rRNA gene-based polymerase chain reaction (PCR). All the confirmed isolates were screened for six toxin genes, namely; cpa, cpb, etx, cpi, cpb2, and cpe using a multiplex PCR.

Results: The PCR amplification of 16S rRNA gene revealed that out of 177 samples collected, 125 (70.62%) were found positive for C. perfringens, of which 110 (72.36%) were from sheep and 15 (60%) were from goats. The highest prevalence of C. perfringens toxinotype D was observed in lambs (56.16%) and kids (46.16%) followed by 3.84% in adult sheep while it was absent in samples obtained from adult goats. The multiplex PCR revealed that 67 (60.90%) isolates from sheep and 8 (53.33%) isolates from goats belonged to toxinotype A, while 43 (39.09%) isolates from sheep and 7 (46.66%) isolates from goats were detected as toxinotype D. None of the isolates was found to be toxinotype B, C, or E. All the C. perfringens toxinotype A isolates from sheep were negative for both cpb2 and cpe genes, however, 27.90% toxinotype D isolates from sheep carried cpb2 gene, and 6.97% possessed cpe gene. In contrast, 12.50% C. perfringens toxinotype A isolates from goats harbored cpb2 and cpe genes while 14.28% isolates belonging to toxinotype D carried cpb2 and cpe genes, respectively.

Conclusion: The high prevalence of C. perfringens was observed, even in day-old lambs. The toxinotypes A and D are prevalent in both sheep and goats. The severity of disease and mortality may be associated with the presence of minor toxins in both the detected toxinotypes.

Keywords: Clostridium perfringens, enterotoxemia, multiplex polymerase chain reaction, toxinotype, 16S rRNA.

Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea)

Research (Published online: 21-12-2017)

13. Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea)

Sakil Ahmed, Aamir Sohail, Sabiha Khatoon, Shabnam Khan and Mohammad Khalid Saifullah

Veterinary World, 10(12): 1493-1500

ABSTRACT

Aim: Aim of the present study was to carry out the partial purification and biochemical characterization of glutathione S-transferase (GST) from the somatic tissue of ruminal amphistome parasite, Gastrothylax crumenifer (Gc) infecting Indian water buffalo (Bubalus bubalis).

Materials and Methods: The crude somatic homogenate of Gc was subjected to progressive ammonium sulfate precipitation followed by size exclusion chromatography in a Sephacryl S 100-HR column. The partially purified GST was assayed spectrophotometrically, and the corresponding enzyme activity was also recorded in polyacrylamide gel. GST isolated from the amphistome parasite was also exposed to variable changes in temperature and the pH gradient of the assay mixture.

Results: The precipitated amphistome GST molecules showed maximum activity in the sixth elution fraction. The GST subunit appeared as a single band in the reducing polyacrylamide gel electrophoresis with an apparent molecular weight of 26 kDa. The GST proteins were found to be fairly stable up to 37°C, beyond this the activity got heavily impaired. Further, the GST obtained showed a pH optima of 7.5.

Conclusion: Present findings showed that GST from Gc could be conveniently purified using gel filtration chromatography. The purified enzyme showed maximum stability and activity at 4°C.

Keywords: Bubalus bubalis, Gastrothylax crumenifer, glutathione S-transferase, purification, somatic tissue.

Seasonal influence on sperm parameters, scrotal measurements, and serum testosterone in Ouled Djellal breed rams in Algeria

Research (Published online: 21-12-2017)

12. Seasonal influence on sperm parameters, scrotal measurements, and serum testosterone in Ouled Djellal breed rams in Algeria

S. Belkadi, B. Safsaf, N. Heleili, M. Tlidjane, L. Belkacem and Y. Oucheriah

Veterinary World, 10(12): 1486-1492

ABSTRACT

Aim: This study was conducted to determine the effect of seasonal variations on testosterone serum concentration, body weight, scrotal circumference, and some sperm parameters in rams living in a semi-arid region of eastern part of Algeria.

Materials and Methods: Blood samples were taken monthly from eight Ouled Djellal rams, aged between 3 and 4 years, in the Technical Institute of Breeding "ITELV" located at Ain M'lila City. Sperm were collected by an electro-ejaculator once a month for 1 year (spring, summer, autumn, and winter: 3 times/season).

Results: Mean values of volume, mass motility, live sperm, and scrotal circumference were higher during spring (p<0.05) with 1.23±0.26 mL, 3.39±1.07, 79.16±15.82%, and 36.29±1.91 cm, respectively; whereas, the sperm concentration was higher during autumn with 1.19±0.56x10⁹ spz/ml compared to 0.46±0.13x10⁹ spz/mL to spring. The season influenced significantly the percentage of abnormal sperm (p<0.05), especially during winter (6.47±2.12%), but had no influence on the weight of rams. Seasonal hormonal activity was high with 4.89±2.06 ng/mL and 3.09±1.35 ng/mL of testosterone in mating seasons (spring and autumn, respectively), knowing that the sexual season is not marked too much in these latitude.

Conclusion: We can conclude that testosterone concentration is strongly correlated with the scrotal circumference and that the season has a significant influence on spermatic parameters, and that despite the large variations in sperm production, the rams can be used throughout the year.

Keywords: scrotal circumference, season, semi-arid area, spermatozoon, testosterone, weight.

The role of staphylococci in subclinical mastitis of cows and lytic phage isolation against to Staphylococcus aureus

Research (Published online: 16-12-2017)

11. The role of staphylococci in subclinical mastitis of cows and lytic phage isolation against to Staphylococcus aureus

Aliye Gulmez Saglam, Mitat Sahin, Elif Celik, Ozgur Celebi, Dogan Akca and Salih Otlu

Veterinary World, 10(12): 1481-1485

ABSTRACT

Aim: This study was conducted to determine the role of Staphylococcus in the formation of subclinical mastitis in cows and to isolate the phage against isolated Staphylococcus aureus strains.

Materials and Methods: In this study, 400 milk cows were screened by California Mastitis Test (CMT) for subclinical mastitis and 235 udders of 96 cows, which were determined to be positive, were evaluated for Staphylococcus. Milk samples were evaluated using conventional and molecular methods. In addition, phage isolation studies were performed against S. aureus strains causing mastitis.

Results: At the result of cultural examination, of 235 milk samples that were found as positive for mastitis by CMT, a total of 117 (49.7%) Staphylococcus spp. were isolated as a distribution of 74 (63.24%) coagulase-positive staphylococci and 43 (36.75%) coagulase-negative staphylococci. Of these isolates, 76 (64.95%) were characterized as S. aureus both conventional and molecular techniques. Lytic bacteriophages against two S. aureus strains which were isolated from mastitic milk samples were obtained from wastewater samples.

Conclusion: The results of this study show that a significant portion of subclinical mastitis was formed by staphylococci. In addition, phage isolation against S. aureus strains isolated can be considered as one of the steps to be applied in the prophylaxis and treatment of such infections.

Keywords: bacteriophage, dairy cow, Staphylococcus aureus, subclinical mastitis.

Prevalence of Theileria annulata in dairy cattle in Nyala, South Darfur State, Sudan

Research (Published online: 15-12-2017)

10. Prevalence of Theileria annulata in dairy cattle in Nyala, South Darfur State, Sudan

Ismail A. Abaker, Diaeldin A. Salih, Lima M. El Haj, Rawia E. Ahmed, Manal M. Osman and Awadia M. Ali

Veterinary World, 10(12): 1475-1480

Aim: This study was conducted in dairy cattle in Nyala, South Darfur State, during the period from June to September 2015, to study the prevalence of bovine tropical theileriosis.

Materials and Methods: Apparently, healthy cattle of different age groups, different breeds, and from both sexes were randomly selected from seven locations. Three age groups of cattle were selected, group one <1 year old, group two 1-3 years old, and group three older than 3 years. These cattle were indigenous and cross (Friesian X zebu). A total of 150 blood samples were collected for blood smears, blood in EDTA tubes, and serum samples as well as ticks infesting cattle. Three diagnostic techniques were used such as blood smear, indirect fluorescent antibody test (IFAT), and polymerase chain reaction (PCR).

Results: Of 150 samples, 11 (7.3%, 95% confidence interval [CI]: 9.1-5.5) were positive for Theileria spp. piroplasms in the blood smears, 70 (46.7%, 95% CI: 35.7-57.7) were positive for Theileria annulata antibodies in the IFAT, and of 100 samples, 39 (39%, 95% CI: 46.6-31.4) were positive for T. annulata using PCR. The prevalence of T. annulata was higher in indigenous breed than cross cattle by the three diagnostic techniques. The highest prevalence of T. annulata was recorded among cattle older than 3 years old. There were three genera and ten species of ticks found feeding on cattle. These were Rhipicephalus evertsi evertsi, Rhipicephalus decoloratus, Rhipicephalus annulatus, Hyalomma dromedrii, Hyalomma impeltatum, Hyalomma rufipes, Hyalomma anatolicum, Hyalomma truncatum, Amblyomma variegatum, and Amblyomma lepidum.

Conclusion: The study concluded that tropical theileriosis is prevalent among dairy cattle in Nyala. H. anatolicum was found in very low numbers, suggesting other ticks may play a role in the transmission of the disease. Molecular characterization of T. annulata is recommended for accurate mapping of the disease and evaluates the magnitude problem of tropical theileriosis in South Darfur region.

Keywords: indirect fluorescent antibody test, polymerase chain reaction, South Darfur, Sudan, Theileria annulata, ticks.

Prevalence of carrier state theileriosis in lactating cows

Research (Published online: 14-12-2017)

9. Prevalence of carrier state theileriosis in lactating cows

Niranjana Sahoo, Bikash Kumar Behera, Hemant Kumar Khuntia and Manojita Dash

Veterinary World, 10(12): 1471-1474

ABSTRACT

Aim: The objective of this study was to examine the carrier status of theileriosis among apparently healthy cross-bred jersey cattle population of Odisha using conventional blood smear examination and polymerase chain reaction (PCR).

Materials and Methods: A total of 34 apparently healthy cross-bred Jersey lactating cows were considered in this study. Blood samples were subjected to microscopic examination after staining with Giemsa stain and PCR based molecular diagnosis using two sets of primer, i.e., N516/N517 and TorF1/TorF2 specific for Theileria annulata and Theileria orientalis, respectively.

Results: Examination of blood samples revealed presence of theileria parasites to a magnitude of 20.59% for T. annulata, 8.82% for T. orientalis, and 2.94% for both.

Conclusion: Molecular diagnosis was found to be much more sensitive than conventional method for diagnosis of theileriosis. T. annulata was found to be the predominant species affecting the exotic cattle. T. orientalis was detected in apparently healthy cows.

Keywords: bovine theileriosis, carrier state, polymerase chain reaction, Theileria annulata, Theileria orientalis.