Genetic variability and discrimination of low doses of Toxocara spp. from public areas soil inferred by loop-mediated isothermal amplification assay as a field-friendly molecular tool

Research (Published online: 27-12-2016)

23. Genetic variability and discrimination of low doses of Toxocara spp. from public areas soil inferred by loop-mediated isothermal amplification assay as a field-friendly molecular tool - Maryam Ozlati, Adel Spotin, Abbas Shahbazi, Mahmoud Mahami-Oskouei, Teimour Hazratian, Mohammad Adibpor, Ehsan Ahmadpour, Afsaneh Dolatkhah and Paria Khoshakhlagh

Veterinary World, 9(12): 1471-1477

   doi: 10.14202/vetworld.2016.1471-1477

Abstract

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Aim: One of the main diagnostic problems of conventional polymerase chain reaction (PCR) is indiscrimination of low parasitic loads in soil samples. The aim of this study is to determine the genetic diversity and identification of Toxocara spp. from public areas soil inferred by loop-mediated isothermal amplification (LAMP) assay.

Materials and Methods: A total of 180 soil samples were collected from various streets and public parks of northwest Iran. The DNA of recovered Toxocara eggs were extracted and amplified by PCR and LAMP following ZnSO4 flotation technique. The amplicons of internal transcribed spacer-2 gene were sequenced to reveal the heterogeneity traits of Toxocara spp. In addition, Toxocara canis sequences of southwest Iran were directly retrieved to compare gene flow between two distinct populations.

Results: Toxocara spp. eggs were found in 57, 14 and 77 of soil samples using the microscopy, PCR and LAMP (detection limit 1-3 eggs/200 g soil), respectively. 7.7% of isolates were identified as T. canis by PCR method, while LAMP was able to detect 27.2%, 15.5% and 12.2% as Toxocara cati, T. canis and mixed infections, respectively. The kappa coefficient between LAMP and microscopy indicated a strong agreement (0.765) but indicated a faint agreement among LAMP-PCR (0.203) and PCR-microscopy (0.308) methods. A pairwise fixation index (Fst) as a degree of gene flow was generally low (0.02156) among Toxocara populations of northwest and southwest Iran.

Conclusions: The statistically significant Fst value indicates that the T. canis populations are not genetically well differentiated between northwest and southwest Iran. This shows that here is possibly an epidemiological drift due to the transfer of alleles. The LAMP assay because of its shorter reaction time, more sensitivity, and simultaneous detection of environmental contamination to be appears as valuable field diagnosis compared to PCR. Therefore, the detection of low Toxocara spp. loads from public area soils will help to expand epidemiological understanding of toxocariasis and establishing preventive strategies in resource-limited endemic of Iran.

Keywords: gene flow, Iran, loop-mediated isothermal amplification, polymerase chain reaction, soil, Toxocara spp.

Estrus induction and fertility response following different treatment protocols in Murrah buffaloes under field conditions

Research (Published online: 22-12-2016)

22. Estrus induction and fertility response following different treatment protocols in Murrah buffaloes under field conditions - L. Kumar, J. B. Phogat, A. K. Pandey, S. K. Phulia, S. Kumar and J. Dalal

Veterinary World, 9(12): 1466-1470

   doi: 10.14202/vetworld.2016.1466-1470

Abstract

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Aim: The aim of this study was to evaluate the efficacy of three different treatment protocols for estrus induction and conception rate in postpartum anestrus buffaloes during breeding season under field conditions.

Materials and Methods: The 47 postpartum anestrus buffaloes of the 2nd to 6th parity were divided into three groups. Group 1 (n=16): Buffaloes received cosynch treatment, that is, buserelin acetate 10 μg on day 0 and 9, cloprostenol 500 μg on day 7 followed by fixed-time artificial insemination (FTAI) at the time of second buserelin acetate and 24 h later. Group 2 (n=15): Buffaloes received norgestomet ear implant subcutaneously for 9 days, estradiol benzoate 2 mg on the day of implant insertion (day 0), pregnant mare serum gonadotropin (PMSG) 400 IU and cloprostenol 500 μg on day 9 followed by AI at 48 and 72 h after implant removal. Group 3 (Cosynch-plus, n=16): Buffaloes received Cosynch protocol as per Group 1 except an additional injection of PMSG 400 IU (i.m.) was given 3 days before the start of protocol and FTAI done at the same time of Group 1. Pregnancy diagnosis was performed after 45 days of AI.

Results: The estrus induction response following the treatment was 81.3%, 100%, and 93.7% in Group 1, 2, and 3, respectively. The buffaloes of Group 1, 2, and 3 expressed intense (38.4%, 60% and 46.6%, respectively) and moderate estrus (46.1%, 26.6%, and 40%, respectively). The conception rates in Group 1, 2, and 3, at FTAI and overall including subsequent estrus were 37.5% and 62.5%, 53.3%, and 66.6%, 56.3%, and 75%, respectively.

Conclusion: All the three treatment protocols can be effectively used for induction of estrus with acceptable conception rate in postpartum anestrus buffaloes during breeding season under field conditions. However, Cosynch-plus (similar to Cosynch protocol except addition of PMSG, 400 IU 3 days before the start of first buserelin acetate administration) protocol results comparatively better pregnancy rate.

Keywords: anestrus, Cosynch-plus, estrus induction, norgestomet.

Survey on Sarcocystis in bovine carcasses slaughtered at the municipal abattoir of El-Kharga, Egypt

Research (Published online: 21-12-2016)

21. Survey on Sarcocystis in bovine carcasses slaughtered at the municipal abattoir of El-Kharga, Egypt - Ali Meawad Ahmed, Nagwa Thabet Elshraway and Ahmed Ibrahim Youssef

Veterinary World, 9(12): 1461-1465

   doi: 10.14202/vetworld.2016.1461-1465

Abstract

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Aim: The main objectives of this study were to determine the incidence of Sarcocystis sp. infection in cattle and buffalo carcasses slaughtered at El-Kharga abattoir, New Valley Governorate, Egypt.

Materials and Methods: The slaughtered animals were daily inspected for Sarcocystis macrocysts through a year (2015). Macroscopic Sarcocystis was detected from a total of 2120 cattle and buffalo carcasses. In addition, 100 meat samples were collected from female cattle and buffalo (50 each) and were examined microscopically for sarcocystosis.

Results: The overall incidence of Sarcocystis macrocyst among bovine carcasses was 159/2120 (7.5%). Total incidence in cattle was 149/2000 (7.45%), whereas it was 10/120 (8.33%) in buffalo carcasses. Concerning gender, the overall prevalence of Sarcocystis infection was 127/1790 (7.09%) in male and 32/330 (9.69%) in females bovine carcasses. The highest detection rate of Sarcocystis lesions was from the esophagus (76.3%) followed by throat muscles (35.3%), tongue (33.8%), and diaphragm muscles (18.71%). Macrocysts from cattle were identified to Sarcocystis hirsuta, whereas Sarcocystis fusiformis was identified from buffalo carcasses. By microscopic examination, 18 (36%) of 50 female cattle carcasses harbor Sarcocystis sp., whereas 11 (22%) of buffalo carcasses were harbored Sarcocystis microcysts.

Conclusion: A high incidence of Sarcocystis infection was detected among slaughtered bovines in El-Kharga abattoir, Egypt. Sarcocystis macrocysts were a higher incidence in female elder animals macrocysts were identified to S. hirsuta in cattle and S. fusiformis in buffaloes. Sarcocystosis constitute a major cause of economic losses at El-Kharga abattoir. Beef meat may carry health risks to consumers.

Keywords: bovine, meat, Sarcocystis fusiformis, Sarcocystis hirsuta, Sarcocystis.

Multiple infections of Anaplasma platys variants in Philippine dogs

Research (Published online: 20-12-2016)

20. Multiple infections of Anaplasma platys variants in Philippine dogs - Adrian Patalinghug Ybañez, Rochelle Haidee Daclan Ybañez, Naoaki Yokoyama and Hisashi Inokuma

Veterinary World, 9(12): 1456-1460

   doi: 10.14202/vetworld.2016.1456-1460

Abstract

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Aim: Anaplasma platys, the causative agent of infectious canine cyclic thrombocytopenia, is a tick-borne pathogen that also has been implicated as potentially zoonotic. To provide molecular evidence on the multiple infections of A. platys variants in Philippine dogs.

Materials and Methods: DNA fragments of A. platys from infected dogs in the Philippines were molecularly characterized. For screening, 25 dogs suspected to have canine anaplasmosis were tested using a 16S rRNA-based nested polymerase chain reaction (PCR). Infection was confirmed by sequencing of positive amplicons. Second round PCR targeting a longer 16S rRNA fragment was subsequently performed on the first round PCR amplicons of the positive samples. Further characterization using the heat-shock operon (groEL) gene was also performed on the A. platys-positive samples.

Results: 10 16S rRNA sequences were obtained and found 99.6-100% identical to each other and 99.6-99.7% identical to the closest registered A. platys sequences. On the other hand, 36 groEL clone sequences were obtained and found to be 85.1-99.8% identical with each other and 85.0-88.9% identical to the closest previously registered A. platys sequences. Four dogs were found coinfected with 2-3 groEL variant sequences. Phylogenetic analyses suggest that the detected A. platys in the Philippines may represent unique variants.

Conclusion: A. platys variants were detected in Philippine dogs. Coinfection of different A. platys variants in dogs was also demonstrated. The present study may indicate the potential genetic diversity of A. platys in the country.

Keywords: Anaplasma platys, dogs, Philippines, variants.

Epidural analgesia in cattle, buffalo, and camels

Review (Published online: 19-12-2016)

19. Epidural analgesia in cattle, buffalo, and camels - Zuhair Bani Ismail

Veterinary World, 9(12): 1450-1455

   doi: 10.14202/vetworld.2016.1450-1455

Abstract

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Epidural analgesia is commonly used in large animals. It is an easy, cheap, and effective technique used to prevent or control pain during surgeries involving the tail, anus, vulva, perineum, caudal udder, scrotum, and upper hind limbs. The objectives of this article were to comprehensively review and summarize all scientific data available in the literature on new techniques and drugs or drug combinations used for epidural anesthesia in cattle, camel, and buffalo. Only articles published between 2006 and 2016 were included in the review. The most common sites for epidural administration in cattle, camels, and buffalos were the sacrococcygeal intervertebral space (S5-Co1) and first intercoccygeal intervertebral space (Co1-Co2). The most frequently used drugs and dosages were lidocaine (0.22-0.5 mg/kg), bupivacaine (0.125 mg/kg), ropivacaine (0.11 mg/kg), xylazine (0.05 mg/kg), medetomidine (15 μg/kg), romifidine (30-50 μg/kg), ketamine (0.3-2.5 mg/kg), tramadol (1 mg/kg), and neostigmine (10 μg/kg), and the clinical applications, clinical effects, recommendations, and side effects were discussed.

Keywords: buffalo, camel, cattle, epidural analgesia, side effects.

Detection, identification, and differentiation of sheep pox virus and goat pox virus from clinical cases in Giza Governorate, Egypt

Research (Published online: 18-12-2016)

18. Detection, identification, and differentiation of sheep pox virus and goat pox virus from clinical cases in Giza Governorate, Egypt - M. A. Mahmoud and M. H. Khafagi

Veterinary World, 9(12): 1445-1449

   doi: 10.14202/vetworld.2016.1445-1449

Abstract

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Aim: To isolate, identify, and differentiate Capripoxviruses (CaPV) (sheep pox virus and goat pox virus) infections by egg inoculation, transmission electron microscopy (TEM), and 30 kDa RNA polymerase subunit gene-based polymerase chain reaction (PCR) (RPO30) in clinically affected animals in Hawamdia township of Giza Governorate, Egypt.

Materials and Methods: A total of 37 scab samples were collected from clinically suspected field cases of sheep pox and goat pox. These samples were collected during (2014-2015) during different outbreaks of sheep pox and goat pox from Hawamdia township of Giza Governorate, Egypt. The samples were subjected to egg inoculation, TEM, and (RPO30) gene-based PCR. By using the egg inoculation: Previously prepared 37 scab samples (n=23 sheep and n=14 goats) were inoculated on the chorioallantoic membrane of specific pathogen free (SPF) embryonated chicken eggs (12 days old age). In the presence of the suitable percentage of humidity and candling, the inoculated eggs were incubated at 37°C. By using the TEM: Samples showed positive pock lesions on the chorioallantoic membranes, were fixed in glutaraldehyde, then processed and sectioned for TEM. Using the (RPO30) gene-based PCR assay, 30 of positive samples after egg inoculation (n=19 sheep and n=11 goats) were screened.

Results: Using the egg inoculation, a characteristic pock lesions for poxviruses were seen in 30/37 (n=19 sheep and n=11 goats) (81.08%). Using the TEM, examination of the positive samples after egg inoculation revealed positive result in 23/30 (n=15 sheep and n=8 goats) (76.66%). The positive results represented by the presence of negatively stained oval-shape virus particles. Using the (RPO30) gene-based PCR assay, out of 30 total of positive samples after egg inoculation (n=19 sheep and n=11 goats) were screened, 27 (90%) samples (n=17 sheep and n=10 goats) were positive. The given band sizes of sheep and goats were 172 and 152 bp, respectively.

Conclusion: PCR assay depended on RPO30 gene can be used lonely for the detection, identification, and differentiation of CaPVs. RPO30 gene-based PCR assay in combination with gene sequencing helps in molecular epidemiological studies of CaPV infection.

Keywords: Capripoxvirus, polymerase chain reaction, goat pox, isolation, RPO30 gene, sheep pox, transmission electron microscope.

Evaluation of sperm recovered after slaughter from cauda epididymides of red Sokoto bucks

Research (Published online: 18-12-2016)

17. Evaluation of sperm recovered after slaughter from cauda epididymides of red Sokoto bucks - A. H. Abu, A. I. Kisani and T. Ahemen

Veterinary World, 9(12): 1440-1444

   doi: 10.14202/vetworld.2016.1440-1444

Abstract

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Aim: Viable spermatozoa could be recovered from the cauda epididymides for the purpose of preservation of genetic material of male animals with desirable traits and for use in reproductive biotechnology. The aim of this study was to determine the effect of storage time on testicular and epididymal biometry, sperm reserves and epididymal sperm characteristics of red Sokoto bucks post mortem.

Materials and Methods: Testes-epididymides were collected immediately after slaughter of mature red Sokoto bucks and transported in ice chest to the laboratory. The samples were either processed immediately or stored at 4°C in refrigerator for 24, 48 h and then processed. The testes and epididymides were measured and weighed. Sperm motility, concentration, livability, morphology, intact acrosome, and sperm reserves from different treatment groups including control were evaluated and means (±standard error of mean) were recorded.

Results: There was no significant difference (p>0.05) in the testicular and epididymal dimensions determined between the means of the groups. Percent sperm motility and viability decreased significantly (p<0.05) after 24 h from 69.00±0.46 and 71.27±0.50% to 50.60±0.48 and 60.47±0.70% at 48 h, respectively. Significant decreases (p<0.05) in epididymal sperm concentration and intact acrosome from 2.86±0.08 and 92.87±0.39 at 0 to 24 h of storage, respectively, were observed. Gonadal sperm reserves and sperm reserves in the cauda epididymides decreased significantly (p<0.05) from 0 to 48 h of cold storage. However, no significant decrease (p>0.05) in caput and corpus sperm reserves was observed from 0 to 48 h of storage.

Conclusion: The results of this study suggest that spermatozoa recovered from the epididymides of red Sokoto bucks were viable after storage for up to 48 h. Furthermore, this finding offers some hope that epididymal sperm recovered post-mortem can be used in assisted reproductive technologies.

Keywords: epididymides, goat bucks, spermatozoa, testes.