Validation of real-time reverse transcription polymerase chain reaction to detect virus titer and thermostability of Newcastle disease live virus vaccine

Research (Published online: 20-11-2018)

12. Validation of real-time reverse transcription polymerase chain reaction to detect virus titer and thermostability of Newcastle disease live virus vaccine

Pangkaj Kumar Dhar, Avijit Dutta, Avijit Das, Mohammad Shah Jalal, Himel Barua and Paritosh Kumar Biswas

Veterinary World, 11(11): 1597-1603

ABSTRACT

Background and Aim: Newcastle disease is one of the most common diseases affecting poultry in Bangladesh. The disease can cause up to 100% mortality but is preventable if birds are timely and properly vaccinated with a vaccine of standard virus titer. Different live vaccines are commercially available in the country - most, if not all, are produced using lentogenic strains of the virus with variable virulence. One of the disadvantages of these vaccines is that they are not stable at high environmental temperature, and therefore, a proper cold chain must be maintained during transportation and storage. Information on how long these vaccine viruses can withstand environmental temperature, which is near the vicinity of 37°C in the summer season in Bangladesh, is scanty. The aim of this research was to measure the effect of temperature on virus titer of live ND virus vaccines and to develop a real-time reverse transcription polymerase chain reaction (rRT-PCR) standard curve to indirectly determine hemagglutination (HA) titer of virus by this highly sensitive method.

Materials and Methods: In this study, thermostability of five commercial live vaccines containing LaSota, F, Clone 30, and B1 type LaSota strains was observed for up to 35 days keeping them at 37°C. From the most thermostability yielding sample, two rRT-PCR standard curves were developed: (1) By plotting the cycle threshold (CT) values as obtained from 10-fold serial dilutions up to 10-3 against their corresponding log (to the base 10) dilutions and (2) by plotting the CT values obtained from serial HA dilutions up to 2-4 against their corresponding HA titer dilutions. The PCR efficiencies based on which the graphs were fitted were also evaluated.

Results: The vaccine from the LaSota strain withstood 37°C for 35 days with a gradual declination of HA titer over time, and this vaccine also had the highest initial HA titer, which was 211. The vaccine made from F strain was inactivated quickly, and it had the lowest HA titer at the beginning of the study. The first standard curve developed can be used to assess the level of virus titer in a diluted sample compared with the titer in the original undiluted vaccine preparation by plotting the CT value obtained from the dilution by rRT-PCR. The second standard curve can be used to calculate the HA titer of a vaccine dilution by plotting the CT value as obtained from the dilution by rRT-PCR.

Conclusion: The regression equations for the first and second graphs were y=–3.535x+14.365 and y=–1.081x+13.703, respectively, suggesting that, for every 3.53 cycles, the PCR product would have increased 10 times and 2 times for every 1.08 cycles, respectively, indicating nearly (but not exactly) 100% PCR efficiency.

Keywords: LaSota strain, Newcastle disease, real-time reverse transcription polymerase chain reaction, thermostability, vaccine, validation.

Effect of feeding rice based distillers dried grains with solubles and gluten meal on nutrient transporter genes and immunity in broiler chickens

Research (Published online: 20-11-2018)

11. Effect of feeding rice based distillers dried grains with solubles and gluten meal on nutrient transporter genes and immunity in broiler chickens

Om Prakash Dinani, Pramod Kumar Tyagi, Asit Baran Mandal, Praveen Kumar Tyagi, Dukare Sagar Popat and Sita Prasad Tiwari

Veterinary World, 11(11): 1592-1596

Aim: The aim of this study was to investigate the effect of feeding rice based distillers dried grains with solubles (rDDGS) and gluten meal on nutrient transporter genes and immunity in broiler chickens.

Materials and Methods: A 2×3 factorial design resulted in six experimental diets, namely T1 (no rDDGS/rice gluten meal [RGM]/enzyme), T2 (no rDDGS/RGM, with multienzymes), T3 (12.5% rDDGS, 15% RGM, no enzyme), T4 (12.5% rDDGS, 15% RGM, with protease enzyme), T5 (10% rDDGS, 12.5% RGM, no enzyme), and T6 (10% rDDGS, 12.5% RGM, with protease enzyme). Each treatment was allocated five replicates of chicks, with eight birds in each. Nutrient transporter genes such as Mucin (MUC 2), excitatory amino acid transporter 3 (EAAT3), and peptide transporter (PepT1) and immunity were estimated using standard procedures.

Results: Feeding rDDGS and RGM combination improved humoral immunity, while cell-mediated immunity did not show any significant (p>0.05) effect on broiler chickens. MUC and PepT1 genes showed significantly (p<0.01) decreased relative fold expression in 12.5% rDDGS +15% RGM combination, while EAAT3 gene showed significantly (p<0.01) decreased relative fold expression in both rDDGS and RGM combination levels.

Conclusion: Thus, it may be concluded that feeding rDDGS and RGM combination improved humoral immunity but had an adverse effect on nutrient transporter gene in broiler chickens.

Keywords: broiler, immunity, nutrient transporter genes, protease, rice distillers dried grains with solubles, rice gluten meal.

Detection of gastrointestinal parasites in small-scale poultry layer farms in Leyte, Philippines

Research (Published online: 19-11-2018)

10. Detection of gastrointestinal parasites in small-scale poultry layer farms in Leyte, Philippines

Rochelle Haidee D. Ybañez, Kurt Jimwell G. Resuelo, Ara Patrice M. Kintanar and Adrian P. Ybañez

Veterinary World, 11(11): 1587-1591

ABSTRACT

Background: Gastrointestinal (GIT) parasites can affect poultry productivity by compromising its health. It is well studied in other countries, but the documented reports in the Philippines have been limited.

Aim: The aim of the present study was to evaluate the presence of GIT parasites in selected small-scale poultry layer farms in Leyte, Philippines.

Materials and Methods: A total of 243 stool samples from eight small-scale poultry layer farms in Leyte, Philippines, were examined for GIT parasites using floatation and sedimentation technique. Profile parameters were also obtained. Fecal samples were collected and analyzed using floatation and sedimentation techniques. Statistical significance between GIT parasite positivity and profile parameters was determined using Chi-square test.

Results: GIT parasites were detected in 92.2% of the samples (24.7% with single infection, 42.0% with 2-3 parasites, and 25.5% with three or more parasites). The common parasites detected were Ascaridia spp. (41.2%), Heterakis spp. (59.3%), Capillaria spp. (10.7%), Eimeria spp. (43.2%), and Strongyloides spp. (74.1%). Some profile parameters, including farm location, years in business, number of workers, nearby water system, the practice of fecal cleaning, and presence of other animals, were found to be significantly associated with GIT positivity.

Conclusion: GIT parasites were detected in the poultry of small-scale layer farms in selected areas in Leyte, Philippines. This finding calls for the importance of routine GIT parasite monitoring and the implied need for regular deworming or dewormer rotation in the area.

Keywords: gastrointestinal parasites, layer chickens, Leyte, Philippines, small-scale farms.

Molecular detection of genes encoding resistance to tetracycline and determination of plasmid-mediated resistance to quinolones in avian pathogenic Escherichia coli in Sukabumi, Indonesia

Research (Published online: 18-11-2018)

9. Molecular detection of genes encoding resistance to tetracycline and determination of plasmid-mediated resistance to quinolones in avian pathogenic Escherichia coli in Sukabumi, Indonesia

Ryan Septa Kurnia, Agustin Indrawati, Ni Luh Putu Ika Mayasari and Adin Priadi

Veterinary World, 11(11): 1581-1586

ABSTRACT

Aim: This study aimed to identify genes encoding resistance to tetracycline (TE) and plasmid-mediated resistance to quinolones in Escherichia coli isolates from clinical cases of avian colibacillosis in Sukabumi, Indonesia.

Materials and Methods: A total of 25 E. coli archive isolates were collected in 2013-2017 from clinical cases of avian colibacillosis in Sukabumi, Indonesia. All isolates were tested for TE and quinolone resistance using the disk diffusion method. TE -resistant E. coli isolates were screened for the presence of tet(A) and tet(B) genes by single polymerase chain reaction (PCR). The qnr(A), qnr(B), and qnr(S) genes were detected by multiplex PCR in quinolone-resistant E. coli isolates.

Results: Result of this study shows that 19 of 25 (76%) E. coli isolates are resistant to oxytetracycline and 64% are resistant to TE; among them, 63.2% and 31.5% were positive tet(A) and tet(B), respectively. 13 out of 25 (52%) are resistant to ciprofloxacin and 36% are resistant to enrofloxacin either norfloxacin; among them, 61.6% were positive qnr(A), 7.7% were positive qnr(B), 23% were positive qnr(S), and 7.7% were positive both of qnr(A) and qnr(S).

Conclusion: This study shows that a few pathogens of E. coli are resistant to TE and quinolone. The frequency of tet and qnr genes that are responsible for this resistance among avian pathogenic E. coli isolates in Sukabumi, Indonesia, was high.

Keywords: Antibiotic, colibacillosis, Escherichia coli, genes, resistance.

Corynebacterium pseudotuberculosis mastitis in Egyptian dairy goats

Research (Published online: 13-11-2018)

8. Corynebacterium pseudotuberculosis mastitis in Egyptian dairy goats

A. M. Nabih, Hany A. Hussein, Safaa A. El-Wakeel, Khaled A. Abd El-Razik and A. M. Gomaa

Veterinary World, 11(11): 1574-1580

ABSTRACT

Background and Aim: Mastitis is an important threat facing goat milk industry and is the most common cause of culling. Efficient control of mastitis, based on efficient diagnosis of diseased animals, would improve milk production and reproductive efficiency. In subclinical mastitis (SCM), infected goats demonstrate neither udder symptoms nor abnormal milk. Corynebacterium pseudotuberculosis is an infectious causative agent of mastitis, mostly results as an extension of infection from the supramammary lymph node, and causes financial losses in the goat industry. This study aimed to estimate the prevalence of SCM with emphasis on C. pseudotuberculosis mastitis in Egyptian dairy goats in the selected farms.

Materials and Methods: A total of 336 half milk samples were collected from 177 dairy goats of various crossbreeds, in mid-to-late lactation period, after clinical examination. All samples were examined bacteriologically, while somatic cell count (SCC) was determined only in 180 half milk samples of the clinically healthy milk samples. The isolated and identified C. pseudotuberculosis was examined for evidence of virulence genes (Phospholipase D [pld] and β-subunit of RNA polymerase [rpoB]) by polymerase chain reaction (PCR).

Results: The prevalence of clinical mastitis was 30.5%, while 69.5% of animals were apparently healthy and secreted milk was normal. Of those 180 clinically healthy half milk samples, 96 milk samples (53.33%) showed SCM as detected by SCC (SCC ≥1,000,000 cells/ml). Coagulase-negative staphylococci were the most prevalent bacteria (41.96%), then Staphylococcus aureus (37.5%) and C. pseudotuberculosis (7.14%). Molecular diagnosis of virulence genes revealed evidence of pld gene in 16 isolates (66.66%), and rpoB gene in 6 samples (25%) of the 24 bacteriologically isolated C. pseudotuberculosis. Here, we describe, for the 1st time, isolation and identification of C. pseudotuberculosis from milk of does suffering from SCM in Egypt.

Conclusion: C. pseudotuberculosis must be considered for routine bacteriological examination of milk from dairy goats, particularly herds with a history of caseous lymphadenitis. Pld gene-based PCR is more reliable than rpoB gene-based ones for the diagnosis of C. pseudotuberculosis.

Keywords: bacteriological investigation, caprine, Corynebacterium pseudotuberculosis, mastitis, phospholipase D, β-subunit of RNA polymerase.

Prevalence of coccidian infection in rabbit farms in North Algeria

Research (Published online: 12-11-2018)

7. Prevalence of coccidian infection in rabbit farms in North Algeria

Samia Maziz-Bettahar, Miriem Aissi, Hacina Ainbaziz, Mohamed Sadek Bachene, Safia Zenia and Fairouz Ghisani

Veterinary World, 11(11): 1569-1573

ABSTRACT

Aim: The aim of the present study was to determine the prevalence and intensity of rabbit coccidiosis (Oryctolagus cuniculus) in North Algeria.

Materials and Methods: During the study, 40 rabbit farms were investigated. The farms are located in the provinces of Tizi Ouzou, Medea, and Djelfa which distributed, respectively, into three regions: East Tell Atlas Mountains, Central Tell Atlas Mountains, and High Plateaus. The number of oocyst per gram of feces (OPG) was determined by McMaster technique, and the Eimeria species were identified using morphological criteria.

Results: In the farms investigated, the prevalence of coccidian infection was estimated to 90% (80.7-99.3%) in rabbits after weaning. The classification of the farms according to their parasite load allowed us to show that 37.5% of the prospective farms have an oocyst excretion between 104 and 5×104 oocysts per gram and 22.5% excrete >5×104 oocysts per gram. Excretion levels by region show that the region of East Tel Atlas Mountains ranks first with 79% of farms with a parasitic load >104 coccidians compared to the regions of Central Tel Atlas Mountains and High Plateaus. In total, eight species of Eimeria were identified from oocyst-positive samples. Mixed infections with four Eimeria species were common. E. magna is the dominant species in comparison with E. media and E. irresidua with respective frequencies of 42.5% and 17.6% and 14.9% (p<0.001). Our results showed that the farms using anticoccidial drugs for their rabbits were low (25%) and the percentage of farms with poor hygienic conditions was 65%. There was a significant association between increased oocysts excretion and control measures of coccidian infection.

Conclusion: The study revealed an overall prevalence of 90% in the three Algerian regions. A strong association was observed between Eimeria infection and hygienic status and preventional chemotherapy.

Keywords: Algeria, coccidiosis, Eimeria, oocysts, prevalence, rabbit.

Epizootiological observations on canine microfilaremia in Gujarat state, India

Research (Published online: 09-11-2018)

6. Epizootiological observations on canine microfilaremia in Gujarat state, India

J. R. Patel, S. Devi, J. P. Varshney and K. M. Jadhav

Veterinary World, 11(11): 1564-1568

ABSTRACT

Aim: The present investigation was conducted to study the prevalence of microfilaremia in dogs in Gujarat.

Materials and Methods: A total of 418 adult dogs aged between 2 and 14 years with signs of weakness and non-specified complaints, presented at TVCC, Deesa (North Gujarat), Nandini Veterinary Hospital, Surat (South Gujarat), and Private Clinics, Ahmedabad (Central Gujarat), were included in the present investigation for studying the prevalence of microfilaremia from July 2016 to May 2017.

Results: A total of 418 dogs were screened, of which 33 were found positive for circulating microfilariae with the prevalence rate of 7.89% in the population. Among microfilaremic dogs, the finding of microfilariae of Dipetalonema (Acanthocheilonema) reconditum was more common (23 cases; 69.69%) than Dirofilaria immitis (10 cases; 30.30%) making their prevalence in the population of 418 dogs as 5.50% and 2.39%, respectively. Breed-wise distribution of microfilaremic dogs revealed that 12 (36.36%), 8 (24.24%), 5 (15.15%), 4 (12.12%), 2 (6.06%), 1 (3.03%), and 1 (3.03%) cases were observed in Pomeranian, non-descript, German Shepherd, Labrador, Great Dane, Lhasa Apso, and Pug dogs, respectively. Of 10 cases of D. immitis, 5, 2, 2, and 1 were observed in Pomeranian, Labrador, non-descript, and Great Dane dogs, respectively. Cases of Dipetalonema reconditum were highest in Pomeranian (7), followed by non-descript (6), German Shepherd (5), Labrador (2), Great Dane (1), Lhasa Apso (1), and Pug (1). Age-wise distribution recorded significantly (p≤0.01) higher number of cases in adult dogs (4-14 years) for D. immitis (30.30%) and D. reconditum (39.39%). Sex-wise distribution of microfilaremic dogs showed that male (22/33, 66.66%) was more predisposed to microfilaremia rather than females (11/33, 33.34%). It is apparent from the study that the number of dogs with microfilaremia due to D. reconditum was significantly (p≤0.01) higher than that of D. immitis.

Conclusion: The present study revealed that microfilaremia due to D. immitis and D. reconditum is prevalent in the state of Gujarat. The infection with D. immitis was associated with severe lung and cardiac pathological manifestations.

Keywords: Dipetalonema reconditum, Dirofilaria immitis, dogs, microfilariae, modified Knott's technique, prevalence.

Risk factors of a viral nervous necrosis disease in grouper (Epinephelus spp.) cultured in Bintan district, Indonesia

Research (Published online: 09-11-2018)

5. Risk factors of a viral nervous necrosis disease in grouper (Epinephelus spp.) cultured in Bintan district, Indonesia

Eka Juniar, Kurniasih Kurniasih and Bambang Sumiarto

Veterinary World, 11(11): 1558-1563

ABSTRACT

Aim: The aim of the study was to determine the prevalence and risk factors of viral nervous necrosis (VNN) disease in tiger grouper cultured in the floating net cage in Bintan district, Indonesia.

Materials and Methods: Sampling used multiple stages with ponds as interest units. The selection conducted by systematic random sampling from the entire net cage in Bintan district. The fish samples were selected based on the appearance of clinical signs of infected fish. The risk factors investigated in this study included net cage, technical, and sample fish information, culture, water quality, and feed management. A total of 195 fishes pooled to the 39 net cages and tested using the nested polymerase chain reaction technique to determine the VNN status. The brain and eye processed for histopathology.

Results: The prevalence rate of VNN on the net cage was 38% (15/39). The risk factors affecting VNN using bivariate analysis was cleaning the net (χ2=9.80; p=0.002), replacement of net cage (χ2=5.20; p=0.0226), and floating net cage technicians knowledge (χ2=4.13; p=0.042). The variables of positive risk factors affecting VNN by multivariate analysis were the juvenile source and the level of mid-weather changes (technician experience and dissolved oxygen [DO]). Seven variables associated to the VNN outbreak have detected. The positive multiplier factors were the source of juveniles, mid weather changes, technician experience, and the DO, while the negative factors were salinity, mixed feed, and the low weather changes. Histopathologically, the grouper fish showed the brain, eye, and muscle vacuolization and kidney necrosis.

Conclusion: It proves that the Bintan waters contaminated by VNN had a prevalence rate of 38% from the total sample based on bivariate method, net cleaning, net replacement, and knowledge on the VNN outbreak. Natural infection of VNN in grouper leads to vacuolization of the brain, the eye, and muscles nearby the eye as well the kidney necrosis.

Keywords: Grouper, multivariate analysis, risk factors, viral nervous necrosis.

The role of E3 ubiquitin ligase seven in absentia homolog in the innate immune system: An overview

Review (Published online: 07-11-2018)

4. The role of E3 ubiquitin ligase seven in absentia homolog in the innate immune system: An overview

Ferbian Milas Siswanto, I. Made Jawi and Bambang Hadi Kartiko

Veterinary World, 11(11): 1551-1557

ABSTRACT

The innate immune system has been considered as an ancient system and less important than the adaptive immune system. However, the interest in innate immunity has grown significantly in the past few years marked by the identification of Toll-like receptors, a member of pattern recognition receptors (PRRs). The PRRs are crucial for the identification of self- and non-self-antigen and play a role in the initiation of signaling events that activate the effective immune response. These sensor signals through interweaving signaling cascades which result in the production of interferons and cytokines as the effector of immune system. Ubiquitin and ubiquitin-like modifiers (UBLs) actively mediate the rapid and versatile regulatory processes that initiate the activation of the innate immune system cascade. The seven in absentia homolog (SIAH) is a potent RING finger E3 ubiquitin ligase that is known to involve in several stress responses, including hypoxia, oxidative stress, DNA damage stress, and inflammation. In this review, the role of SIAH will be discussed as an E3 ubiquitin ligase on the regulation of innate immune.

Keywords: E3 ligase, innate immunity, regulation, seven in absentia homolog.

Yields, chemical composition, and antimicrobial activity of two Algerian essential oils against 40 avian multidrug-resistant Escherichia coli strains

Research (Published online: 06-11-2018)

3. Yields, chemical composition, and antimicrobial activity of two Algerian essential oils against 40 avian multidrug-resistant Escherichia coli strains

Narimene Mansouri, Leila Aoun, Nabila Dalichaouche and Douniazed Hadri

Veterinary World, 11(11): 1539-1550

ABSTRACT

Aim: The aim of this study is to investigate, in vitro, a possible antibacterial activity of Algerian essential oils (EOs) of Thyme (Thymus vulgaris L.) and that of Coriander (Coriandrum sativum L.) against multidrug-resistant avian Escherichia coli strains and this in a perspective of their future use as a substitute for antibiotics (ATBs).

Materials and Methods: In addition to the reference strain of E. coli ATCC 25922, 40 strains of avian E. coli have been isolated (24 strains of broilers and 16 of turkeys), their antimicrobial resistance profile was determined by antibiogram tests against 21 ATBs whereupon they were subjected to the action of two Algerian EOs; the EO of Thyme (T. vulgaris L.) and that of Coriander (C. sativum L.), which oils were extracted by hydrodistillation and analyzed by Gas Chromatography coupled to Mass Spectrometry (GC-MS) and this for the determination of their chemical composition. The antibacterial activity, resulting in zones of inhibition, was evaluated by carrying out, in triplicate, aromatograms for both pure EO and that which has been diluted to 15% in Dimethyl Sulfoxide (DMSO), while the minimum inhibitory concentrations (MIC) of the two EOs were highlighted by the method of liquid macrodilution.

Results: Antibiogram performance demonstrated an alarming state of antimicrobial resistance, the multidrug resistance rate was estimated at 100% for the broilers chicken strains and at 81.25% for strains isolated from turkeys, hydrodistillation allowed to obtained EOs with yields estimated at 1.22±0.26% for Thyme EO and 0.23±0.15% for the essence of Coriander, the GC-MS analysis identified 19 main compounds and showed that the majority chemical components were Carvacrol (73.03%) for Thyme volatile oil and Linalool (60.91%) for Coriander EO, aromatograms and the determination of MIC concluded that the EO of Thyme showed a greater antibacterial activity with an average of the zones of inhibition estimated at 26.75±0.426 mm and MIC ranging from 0.07 to 0.93 mg/ml against an average of the inhibition zones evaluated at 17.05±0.383 mm and MICs evaluated between 0.6 and 10 mg/ml for the EO of Coriander.

Conclusion: In aviculture, these results seem to be very promising in the case where we think about the replacement of ATBs by EOs, in vivo studies would be very interesting to confirm or invalidate this hypothesis.

Keywords: Avian Escherichia coli, antimicrobial resistance, essential oils, antibacterial activity, Thymus vulgaris L., Coriandrum sativum L.

Impact of the flour of Jerusalem artichoke on the production of methane and carbon dioxide and growth performance in calves

Research (Published online: 03-11-2018)

2. Impact of the flour of Jerusalem artichoke on the production of methane and carbon dioxide and growth performance in calves

Sintija Jonova, Aija Ilgaza, Inga Grinfelde and Maksims Zolovs

Veterinary World, 11(11): 1532-1538

ABSTRACT

Aim: The aim of the research was to evaluate the growth performance, to measure the amount of methane (CH4) and carbon dioxide (CO2) in calves' rumen, and to compare the obtained results between the control group (CoG) and the experimental group (Pre12) which received the additional supplement of the prebiotic inulin.

Materials and Methods: The research was conducted with ten Holstein Friesian (Bos taurus L.) crossbreed calves with an average age of 33±6 days. Calves were split into two groups: 5 calves that were fed with the control non-supplemented diet (CoG) and 5 calves that were fed with the same diet further supplemented with 12 g of flour of Jerusalem artichoke (Helianthus tuberosus L.) containing 6 g of prebiotic inulin per 0.5 kg of barley flour diet (Pre12). The duration of the experiment was 56 days. CH4 and CO2 were measured using cavity ringdown spectroscopy device Picarro G2508. The weight and samples from calves' rumen were evaluated 3 times during the experimental period - on the 1st, 28th, and 56th days. Samples were obtained by puncturing the calf rumen.

Results: The weight gain (kg) during the whole experimental period was higher in the Pre12 (65.8±6.57) compared to CoG (36.8±7.98) calves (p<0.001). The daily weight gain was also increased in the Pre12 (1.2±0.12) than CoG (0.7±0.14) calves (p<0.001). There was no difference in the levels of CH4 and CO2 produced in the rumen of CoG and Pre12 calves (p>0.05).

Conclusion: The main results showed that the prebiotic inulin can promote weight gain in calves, without affecting the mean concentration of CH4 and CO2 in calves' rumen.

Keywords: calves, carbon dioxide, inulin, methane, weight gain.

The profiling of pre- and post-warming DNA in mouse embryos with microsatellite method

Research (Published online: 02-11-2018)

1. The profiling of pre- and post-warming DNA in mouse embryos with microsatellite method

Widjiati Widjiati, Soeharsono Soeharsono and Yeni Dhamayanti

Veterinary World, 11(11): 1526-1531

ABSTRACT

Aim: This research aimed to identify the deoxyribonucleic acid (DNA) profile and changes of post-warming embryo after being frozen with vitrification method using microsatellite method.

Materials and Methods: This research examined the mouse embryo blastocysts that were divided into four groups: Post-warming living blastocyst, post-warming living blastocyst with half fragmented cell, post-warming dead blastocyst, and pre-freezing living blastocyst. The isolation sample applied phenol-chloroform method. After obtaining polymerase chain reaction results, all the samples of pre-freezing fresh embryo, post-warming living embryo, dead embryo, and degenerated embryo were then examined by single-strand conformation polymorphism (SSCP).

Results: The amplification with D18mit14 primer was 100 bp and 150bp with D18mit87 primer, 150bp with D7mit22, and 300bp with D7mit25. The result of SSCP with D18mit14 primer showed that the blastocysts were fragmented and dead after warming process and formed into two DNA strand fragments, while the fresh embryos which passed freezing process did not form any fragment. D18mit87 primer SSCP indicated different fragments for each treatment. The result of SSCP using D7mit22 formed two different fragments for each treatment. While using D7mit25, the SSCP result formed some different fragments for each sample. Post-warming living embryo had similar ribbon to pre-freezing fresh embryo.

Conclusion: D7mit222, D7mit25, and D18mit87 primers could be used as the aneuploidy marker on mouse embryos that were induced by post-warming process. The profile of living blastocyst, dead blastocyst, and post-warming fragmented blastocyst had different DNA tapes.

Keywords: blastocyst, DNA mutation, single-strand conformation polymorphism, vitrification.

Preliminary field survey on Mycobacterium bovis infection in cattle herds using caudal fold intradermal tuberculin test in two Northeastern States of Nigeria

Research (Published online: 01-11-2018)

9. Preliminary field survey on Mycobacterium bovis infection in cattle herds using caudal fold intradermal tuberculin test in two Northeastern States of Nigeria

Salisu Ibrahim, Bello Abubakar Usman, Danbirni Samaila and Adamu Saleh Saidu

International Journal of One Health, 4: 52-58

ABSTRACT

Aim: A survey was conducted to determine the prevalence of bovine tuberculosis (bTB) in two states of Northeastern Nigeria, namely Bauchi and Gombe States, between February 2010 and November 2014 using caudal fold tuberculin (CFT) skin test.

Materials and Methods: A total of 5489 cattle were screened using single CFT in Bauchi and Gombe States. Of the 5489 cattle, 2116 cattle were from 189 herds in five testing areas in Bauchi State and 3373 cattle in 217 herds from five testing areas from Gombe State.

Results: On the basis of the CFT, herd prevalence obtained was 56.08% in Bauchi and 55.29% in Gombe State, while the individual animal prevalence was 10.96% in Bauchi and 13.73% obtained in Gombe State. The prevalence based on the testing areas in Bauchi State, Alamari had the highest prevalence with 19.4 % and Disina the lowest with 9.0 %, while in Gombe State, Wakaltu had the highest prevalence with 20.9 % and Poshereng the lowest with 8.0 %. Cows were more likely to have tuberculosis lesions than bulls (p=0.0035) in Bauchi State, but there was no significant difference in Gombe State (p=0.166). However, a statistically significant association (p<0.05) was observed among the cattle age groups with cattle ≥4 years having higher odds for tuberculin reactivity compared to those below the age of 4 years in Bauchi State only.

Conclusion: There is the need to strengthen routine meat inspection and public health awareness programs on the zoonotic nature of bTB among the abattoir workers and the herdsmen.

Keywords: bovine tuberculosis, cattle herds, caudal folds, Northeastern Nigeria, tuberculin test.

Identification and characterization of a novel infectious bursal disease virus from outbreaks in Maharashtra Province of India

Research (Published online: 29-10-2018)

25. Identification and characterization of a novel infectious bursal disease virus from outbreaks in Maharashtra Province of India

Sudhakar P. Awandkar, Prabhakar A. Tembhurne, Jeevan A. Kesharkar, Nitin V. Kurkure, Sandeep P. Chaudhari, Sachin W. Bonde and Vijay C. Ingle

Veterinary World, 11(10): 1516-1525

ABSTRACT

Aim: The study was undertaken to isolate infectious bursal disease virus (IBDV) from clinical cases in broiler and cockerel flocks of Maharashtra state, India, and its molecular epidemiological investigation.

Materials and Methods: The morbid bursal tissues were collected from flocks suspected for IBD. The samples were subjected for virus adaptation in primary chicken embryo fibroblast (CEF) cells followed by confirmation by reverse transcription polymerase chain reaction (RT-PCR) for partial VP2 sequence and phylogenetic analysis.

Results: The isolation of IBDV from field samples took seven blind passages for adaptation in CEF. The cytopathic effects included rounding, aggregation, vacuolation, and detachment of the cells. The RT-PCR showed amplification of 627 bp amplicon specific to the primers for VP2 gene fragment which confirmed successful adaptation and isolation of IBDV using CEF. The nucleotide and deduced amino acids based on phylogeny clustered the current isolate in a distinct clade with classical virulent and antigenic variants. It showed divergence from very virulent (vv) and vaccine strains of Indian origin. The isolate showed unique amino acid substitution at A329V as compared to all other IBDVs. The variation in key amino acids was reported at A222, I242, Q249, Q253, A256, T270, N279, T284, I286, L294, N299, and V329. It shared conserved amino acids at position A222, I242, and Q253 as reported in vvIBDV isolates. However, the amino acids reported at position T270, N279, T284, L294, and N299 are conserved in classic, antigenic variant and attenuated strains of IBDV. The amino acids at positions N279 and T284 indicated that the isolate has key amino acids for cell culture replication.

Conclusion: The IBDV field isolate does not reveal the full nucleotide sequence signature of vvIBDV as well as vaccine strains. Hence, we can conclude that it might not belong to vvIBDVs of Indian origin and the vaccine strain used in the region. This may be suggestive of the evolution of the IBDV in the field due to the coexistence of circulating field strains and live attenuated hot strains, resulting into morbidity and mortality, warranting the need for safer protective vaccines, and implementation of stringent biosecurity measures to minimize loss to farmers.

Keywords: adaptation, chicken embryo fibroblast, epidemiology, isolate infectious bursal disease virus, VP2.

Genetic resistance of eight native Egyptian chicken breeds having chicken B-cell marker 6 gene post-challenge with field strain of Marek's disease-induced tumor virus

Research (Published online: 29-10-2018)

24. Genetic resistance of eight native Egyptian chicken breeds having chicken B-cell marker 6 gene post-challenge with field strain of Marek's disease-induced tumor virus

Hala A. Shaheen, H. A. Hussein, M. M. Elsafty and M. A. Shalaby

Veterinary World, 11(10): 1510-1515

ABSTRACT

Aim: The aim of this work was to detect chicken B-cell marker 6 (ChB6) gene in some native breeds in Egypt and find the relationship between founded genes in these different breeds to determine the resistance of native Egyptian breeds of chicken to Marek's disease (MD).

Materials and Methods: A total of 14 different chicken breeds (30 each) including ten native breeds in addition to SPF Lohmann, High Line, Bovans, and Roodiland were used. Blood samples were collected for the detection of (ChB6,) by polymerase chain reaction (PCR) assay and sequenced to determine the presence or absence of ChB6 gene. Experimental infection was done using local field isolated MD virus (MDV) of 11 (1 day old) unvaccinated chick breeds having no maternal antibodies against MDV. Ten breeds of them carry ChB6 gene, eight breeds were native, and the rest two breeds were SPF Lohmann and High Line in addition to a group of ChB6 gene-lacking breed (Bovans) were infected. Spleen samples were collected from all infected breeds at 20th, 25th, 30th, 35th, and 40th weeks post-infection and tested by PCR assay for the detection of MDV. Furthermore, at 40th week post-infection, tumorized spleen sample of Bovans breed was collected and prepared for examination by transmission electron microscope (TEM) to confirm the presence of MDV.

Results: Our results revealed the positivity of 10 out of 14 breeds (Gimmizah , Sinai, Dandarawi, Fayoumi, Golden Montazah, Matrouh, Beheri, Dokki, SPF Lohmann, and High Line) to the presence of ChB6 gene and resistance to MDV infection, while the Bovans, Mandarah , Inshas and Roodiland breeds lack the ChB6 gene and are susceptible to MDV infection. The collected spleen samples revealed negative for the presence of challenged MDV by PCR in 10 breeds (Gimmizah, Sinai, Dandarawi, Fayoumi, Golden Montazah, Matrouh, Beheri, Dokki, SPF Lohmann, and High Line) and positive for Bovans breed. TEM is used to confirm MDV infection in Bovans group which demonstrated tumors.

Conclusion: The study confirms the relationship between the presence of ChB6 gene in our native breeds and the absence of tumors.

Keywords: ChB6 gene, Egyptian chicken breeds, Marek's disease, polymerase chain reaction, transmission electron microscope.

Bacteriological survey of bulk tank milk from dairy farms in Montero, Santa Cruz, Bolivia

Research (Published online: 26-10-2018)

23. Bacteriological survey of bulk tank milk from dairy farms in Montero, Santa Cruz, Bolivia

Yu Michimuko-Nagahara, Yu Tamura and Masateru Koiwa

Veterinary World, 11(10): 1506-1509

ABSTRACT

Background and Aim: Recently, bacterial surveys for mastitis-causing pathogens in bulk tank milk (BTM) have been conducted in several countries worldwide. However, no such surveys have been reported from Bolivia. Therefore, the present study aimed to estimate the prevalence of mastitis pathogens in BTM from dairy farms in Montero, Santa Cruz, Bolivia.

Materials and Methods: Between July 2016 and August 2017, a total of 43 BTM samples were collected from 3264 cows to determine bulk tank somatic cell counts (BTSCC) and identify mastitis-causing bacteria. BTSCC was classified as follows: = <100×103, 100-500×103, 500-1000×103, and >1000×103 cells/mL.

Results: Mastitis-causing pathogens identified by agar medium cultures included Bacillus spp., coagulase-negative staphylococci (CNS), coliforms, Staphylococcus aureus (SA), streptococci, and other species. The proportions of BTSCC of <100×103, 200-500×103, 500-1000×103, and >1000×103 cells/ml were 0%, 37%, 51%, and 12%, respectively. The proportions of coliforms, streptococci, CNS, Bacillus spp., SA, and others detected in BTM were 33%, 30%, 16%, 7%, 2%, and 16%, respectively.

Conclusion: Although the herd prevalence of contagious mastitis-causing pathogens, such as SA, in BTM was low, increased BTSCC were identified in Montero, Santa Cruz, Bolivia.

Keywords: bacterial survey, Bolivia, bulk tank milk, dairy farm, mastitis.

The effect of Toxoplasma gondii on plasma serotonin concentration in sheep

Research (Published online: 26-10-2018)

22. The effect of Toxoplasma gondii on plasma serotonin concentration in sheep

Annamaria Castello, Giuseppe Bruschetta, Renato Paolo Giunta, Anna Maria Fausta Marino and Alida Maria Ferlazzo

Veterinary World, 11(10): 1500-1505

ABSTRACT

Background and Aim: Toxoplasma gondii is an intracellular parasite that commonly infects warm-blooded animals, including humans. Virtually all species can be infected, but a species-specific variability is evident, in terms of both type and severity of the symptoms encountered. As serotonin (5-hydroxytryptamine [5-HT]) plays an important regulatory role in both physiological and immune responses, the aim of this research was to assess whether toxoplasmosis disease could affect plasma 5-HT concentration and/or hematochemical parameters in a particularly susceptible species to infection as sheep.

Materials and Methods: 5-HT plasma levels were analyzed in platelet-poor plasma fraction by enzyme-linked immunosorbent assay. Blood count and hematochemical parameters were evaluated. Total proteins (TPs), glucose (Glu), and lactate dehydrogenase were determined by a spectrophotometer.

Results: Results showed significantly higher levels in plasma 5-HT, monocytes, and TP and significantly lower levels of Glu, in infected sheep compared to the control group.

Conclusion: Results could support the hypothesis of an effect of toxoplasmosis infection on plasma 5-HT concentrations in sheep. More research is needed to assess the function of 5-HT in the regulation of infected sheep's immune responses.

Keywords: monocytes, plasma, serotonin, serum, sheep, toxoplasmosis.

Evaluation of antioxidant, diuretic, and wound healing effect of Tulkarm honey and its effect on kidney function in rats

Research (Published online: 25-10-2018)

21. Evaluation of antioxidant, diuretic, and wound healing effect of Tulkarm honey and its effect on kidney function in rats

Hamada Imtara, Noori Al-Waili, Meryem Bakour, Wail Al-Waili and Badiaa Lyoussi

Veterinary World, 11(10): 1491-1499

ABSTRACT

Aim: The composition and activity of honey depend on its floral origin. Honey collected from Tulkarm was evaluated for physicochemical property and antioxidant content as well as a diuretic and wound healing activity. Its effect on kidney function was evaluated and compared with furosemide.

Materials and Methods: Honey was collected in Tulkarm, Palestine, and its phenol, flavones, and flavonol content were assessed. The antioxidant activity was determined with the use of colorimetric assays, 1,1-diphenyl-2-picrylhydrazyl, ferric reducing antioxidant power, and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid). Two sets of experiments were conducted. First experiment: 18 rats were used for the evaluation of diuretic activity of honey. The rats received either honey or furosemide. Renal function test, uric acid, and serum and urine electrolytes assay were performed. Second experiment: 18 male mice were used to evaluate the wound healing property of honey. Wounds were created on mice skin and treated daily with honey or Madecassol. Measurements of wounds were performed over a period of 12 days.

Results: The physical and chemical parameters of Tulkarm honey are within the limits of the European legislation and fulfilling the criteria described in the standard codex for honey. It contains antioxidant compounds and shows antioxidant activity. Oral honey increased creatinine clearance and urine volume, sodium, and chloride without causing hypokalemia or affecting blood urea, uric acid, or serum creatinine level. The diuretic activity of furosemide was associated with hypokalemia. Topical honey application enhanced wound closure when compared with the Madecassol application.

Conclusion: The study is the first to report that honey collected from Tulkarm has a considerable diuretic effect without affecting serum electrolytes or kidney function test and exhibits strong antioxidant activity and wound healing property.

Keywords: furosemide, honey, kidney, Madecassol, wounds.

An analysis of common foodborne parasitic zoonoses in slaughtered sheep and cattle in Tehran, Iran, during 2015-2018

Research (Published online: 24-10-2018)

20. An analysis of common foodborne parasitic zoonoses in slaughtered sheep and cattle in Tehran, Iran, during 2015-2018

Ali Pezeshki, Hadi Aminfar and Majid Aminzare

Veterinary World, 11(10): 1486-1490

ABSTRACT

Background and Aim: Cystic echinococcosis, Echinococcus granulosus, and liver flukes, such as Fasciola spp. and Dicrocoelium dendriticum, are important parasitic zoonoses, where they able to cause significant veterinary, medical, and economic problems. The present study was carried out to obtain the updated knowledge on the frequency of hydatidosis, fasciolosis, and dicrocoeliosis in the slaughtered sheep and cattle.

Materials and Methods: Information were collected from meat inspection records using systematically visual inspection, palpation, and incision of the visceral organs in the industrial abattoir in Tehran, the capital of Iran, between February 1, 2015, and January 31, 2018. For an analysis of the data, SPSS version 16 was applied.

Results: The hydatidosis infection in sheep and cattle was 2.48% and 2.25%, respectively. With respect to liver flukes, 0.62% and 0.25% sheep and cattle were infected by Fasciola spp., respectively; furthermore, 2.86% sheep and 0.79% cattle were positive for D. dendriticum.

Conclusion: The findings will provide considerable awareness for the future monitoring and control of these potentially important infections.

Keywords: dicrocoeliasis, fascioliasis, hydatidosis, sheep and cattle, slaughterhouse.