Sequence analysis of the Hex A gene in Jacob sheep from Bulgaria

Research (Published online: 08-01-2021)

7. Sequence analysis of the Hex A gene in Jacob sheep from Bulgaria

Boyko Neov, Jivko Krastanov, Teodora Angelova, Nadezhda Palova, Stayka Laleva and Peter Hristov

Veterinary World, 14(1): 56-60

ABSTRACT

Background and Aim: Jacob sheep are a rare ancient breed of sheep believed to have originated from the Mediterranean area but which are now kept throughout the world. These sheep have recently attracted medical interest due to the observation of a genetic disorder in the breed that can be used as an animal model of Tay–Sachs disease (TSD). This study aims to detect mutations in the Hexosaminidase A gene in Jacob sheep based on sequence analysis of the 284-bp fragment situated between exon 11 and intron 11 of the gene, a target sequence for site-specific mutation. This is the first study that has investigated Jacob sheep in Bulgaria for gene-specific mutations.

Materials and Methods: A total of 20 blood samples were collected from Jacob sheep from the Rhodope Mountains. DNA was isolated from these samples, and a specific 284-bp fragment was amplified. The amplified products were purified using a polymerase chain reaction purification kit and sequenced in both directions.

Results: Target sequences were successfully amplified from all 20 investigated sheep. Sequence analysis did not show the homozygous, recessive, missense (G-to-C transition) mutation at nucleotide position 1330 (G1330→C) in exon 11, demonstrating that all of these sheep were a normal genotype (wild-type).

Conclusion: Jacob sheep are considered a potentially useful animal model in advancing the understanding of pathogenesis and developing potential therapies for orphan diseases, such as those characterized by mutant GM2 gangliosides. The clinical and biochemical features of the Jacob sheep model of TSD represent well the human classical late-infantile form of this disorder, indicating that the model can serve as a possible new research tool for further study of the pathogenesis and treatment of TSD.

Keywords: GM2 gangliosides, Hexosaminidase A gene, human replacement therapy, Jacob sheep, Tay–Sachs disease.

Demographic and clinical characteristics of dogs with centroblastic lymphoma

Research (Published online: 07-01-2021)

6. Demographic and clinical characteristics of dogs with centroblastic lymphoma

Katarzyna Kliczkowska-Klarowicz, Dariusz Jagielski, Michał Czopowicz and Rafał A. Sapierzyński

Veterinary World, 14(1): 49-55

ABSTRACT

Background and Aim: Centroblastic lymphoma (CBL) is the most common morphological type of lymphoma found in dogs; it is usually identified through cytology in veterinary clinical practice. This study aimed to identify the demographic and clinical characteristics of dogs with CBL that was diagnosed with cytology and immunocytochemistry.

Materials and Methods: Dogs with a suspicion of lymphoma were diagnosed by cytology supported by immunocytochemistry with the use of the updated Kiel classification adapted for dogs. During the analyzed time period, 336 lymphomas were diagnosed in dogs, including 171 cases of CBL. Epidemiological and clinical data from the dogs with CBL were provisionally collected.

Results: The epidemiology analysis revealed an increased risk of CBL in Rottweilers, golden retrievers, and Bernese mountain dogs. At admission, most of the dogs displayed generalized lymphadenopathy with spleen and liver enlargement. The most common hematological abnormality was leukocytosis due to neutrophilia. The most common biochemical abnormality was elevated alanine aminotransferase and alkaline phosphatase activities and selective hypoproteinemia due to hypoalbuminemia.

Conclusion: Rottweilers, Bernese mountain dogs, and golden retrievers appear to be overrepresented among dogs with CBL. CBL is usually diagnosed at an advanced clinical stage according to the World Health Organization; however, it is usually accompanied by only minor hematological and biochemical abnormalities.

Keywords: Bernese mountain dogs, clinical stage, cytology, epidemiology, fine-needle biopsy, Golden Retrievers, immunophenotype, lymphoma, Rottweilers.

Role of microorganisms isolated from cows with mastitis in Moscow region in biofilm formation

Research (Published online: 07-01-2021)

5. Role of microorganisms isolated from cows with mastitis in Moscow region in biofilm formation

Pavel Rudenko, Nadezhda Sachivkina, Yury Vatnikov, Sergey Shabunin, Sergey Engashev, Svetlana Kontsevaya, Arfenia Karamyan, Dmitry Bokov, Olga Kuznetsova and Elena Vasilieva

Veterinary World, 14(1): 40-48

ABSTRACT

Background and Aim: Mastitis is one of the most important diseases of cows and the most expensive pathology for the dairy industry. Therefore, this study was conducted to explore the role of microorganisms isolated from cows with mastitis in the formation of biofilms under the conditions of farm biogeocenosis in the Moscow region.

Materials and Methods: Periodic visits to 12 farms in the Moscow region were conducted to explore the microbial profile of the udder of cows with mastitis. During the visits, 103 milk samples from sick animals were collected and examined. Through microbiological analyses, 486 cultures of microorganisms were identified, which are assigned to 11 genera. Mastitis in cows is caused not only by a single pathogen but also by microbial associations, which included two to seven microbial isolates.

Results: It was observed that 309 isolates (63.6%) from the total number of isolated microorganisms could form a biofilm. The ability to form biofilms was most frequently observed in Staphylococcus aureus (18.8%), Escherichia coli (11.9%), and Staphylococcus uberis (11.7%) cultures from the total number of biofilm-forming microbial cultures. Low biofilm-forming ability among the isolated microorganisms was found in lactobacilli, wherein only 20 (22.5%) Lactobacillus strains had the ability to form biofilms. The isolated microorganisms exhibited different sensitivities to antimicrobial agents, which cause difficulty in selecting an antimicrobial agent that would act on all aspects of the parasitocenosis.

Conclusion: A high proportion of microorganisms isolated from cows with mastitis have the ability to form biofilms. The isolated microorganisms exhibited different and highly heterogeneous sensitivity to the action of antimicrobial drugs. This causes difficulty in using these tools for the effective control of mastitis in cows, which is frequently caused by pathogenic associations of microbial biofilms. Therefore, it is important to explore novel and more effective methods to combat this disease.

Keywords: bacterial communities, biofilms, biogeocenosis, cows, mastitis, sensitivity to antibiotics.

Multidrug resistance protein structure of Trypanosoma evansi isolated from buffaloes in Ngawi District, Indonesia: A bioinformatics analysis

Research (Published online: 06-01-2021)

4. Multidrug resistance protein structure of Trypanosoma evansi isolated from buffaloes in Ngawi District, Indonesia: A bioinformatics analysis

Moh. Mirza Nuryady, Raden Wisnu Nurcahyo, Iin Hindun and Diani Fatmawati

Veterinary World, 14(1): 33-39

ABSTRACT

Background and Aim: Trypanosomiasis, also known as surra, is an infectious disease with a wide host spectrum. In Indonesia, this disease is caused by Trypanosoma evansi. Various trypanocidal drugs have been used to treat this pathogen and subsequent disease. Yet, the long-term trypanocidal administration generates drug-resistant T. evansi. Some have identified genetic alterations in T. evansi transporter protein-coding genes that may be responsible for drug resistance. The Multidrug Resistance Protein E (MRPE) gene is a likely candidate gene responsible for the individual resistance. To date, no research has focused on T. evansi MRPE (TevMRPE) in this context. Hence, this research aimed at analyzing and characterizing the TevMRPE gene and protein using a bioinformatics approach.

Materials and Methods: T. evansi was isolated from buffalo suffering from surra in Ngawi Regency, Indonesia. Isolated T. evansi was inoculated and cultured in male mice. The T. evansi genome was isolated from mouse blood with a parasitemia degree as high as 105. A polymerase chain reaction procedure was conducted to amplify the putative MRPE coding gene. The amplicon was sequenced and analyzed using MEGA X, BLAST, and I-tasser softwares.

Results: The putative TevMRPE coding gene showed sequence similarity as high as 99.79% against the MRPE gene from Trypanosoma brucei gambiense. The protein profile and characteristics depicted that the putative TevMRPE protein was related to a family of Adenosine Triphosphate-Binding Cassette (ABC) transporter proteins. This family of transporter proteins plays a crucial role in the resistance toward several medicines.

Conclusion: The obtained gene sequence in this research was identified as the TevMRPE. This gene is homologous to the T. brucei gambiense MRPE gene and possesses ligand active sites for Adenylyl Imidodiphosphate. In addition, MRPE contains enzyme active sites similar to the cystic fibrosis transmembrane conductance regulator. These data suggest that ABC transport proteins, like MRPE, may be necessary to confer trypanocidal drug resistance in T. evansi.

Keywords: bioinformatics, buffaloes, multidrug resistance protein E, protein structure, Surra, Trypanosoma evansi.

Saccharomyces cerevisiae, bentonite, and kaolin as adsorbents for reducing the adverse impacts of mycotoxin contaminated feed on broiler histopathology and hemato-biochemical changes

Research (Published online: 05-01-2021)

3. Saccharomyces cerevisiae, bentonite, and kaolin as adsorbents for reducing the adverse impacts of mycotoxin contaminated feed on broiler histopathology and hemato-biochemical changes

Toochukwu Ejiofor, Anthony Christian Mgbeahuruike, Chiamaka Ojiako, Ashang Micheal Ushie, Emmanuela Ifeoma Nwoko, Ibe Remigius Onoja, Toluwase Dada, Mulunda Mwanza and Magnus Karlsson

Veterinary World, 14(1): 23-32

ABSTRACT

Background and Aim: Saccharomyces cerevisiae, bentonite and kaolin were used to reduce the adverse effects of mold-contaminated diet on broilers. The aim of the study was to evaluate the impact of S. cerevisiae, bentonite, and kaolin in reducing the adverse effects of mold (fungal) contaminated diet on broilers. Specifically, we investigated the histopathological, hematological, and serum biochemical changes associated with broilers fed mold-contaminated diets supplemented with these three adsorbents. We also isolated and identified the common fungal contaminants in the poultry feeds as well as the mycotoxins they produced.

Materials and Methods: Hundred broilers (3-weeks-old) were randomly grouped into five dietary treatments, basal feed (negative control), feed contaminated with mold, mold-contaminated feed+S. cerevisiae, mold-contaminated feed+bentonite, and mold-contaminated feed+kaolin. The fungal contaminants in the feeds were isolated and molecularly identified while the mycotoxins in the feed where analyzed using high-performance liquid chromatography. Blood samples of birds from each group were analyzed for hematology and serum biochemistry. The liver, spleen, kidney, and bursa of Fabricius of the birds were excised and analyzed for histopathological changes.

Results: The most common fungal contaminants in the feeds were Penicillium (33.3%) species, followed by Aspergillus species (22.2%). The mold-contaminated feed had the highest number of fungal contaminants, 55.6%, while the negative control (basal feed group) had none. Total aflatoxin and deoxynivalenol were high in the mold-contaminated feed (53.272 μg/kg and 634.5 μg kg, respectively), but these were reduced by the addition of adsorbents to the feed. The birds fed mold-contaminated feed had significantly (p<0.05) reduced red blood cell count counts, packed cell volume, and hemoglobin but increased white blood cell count compared to the negative control. Liver enzyme activity (alanine transaminase, aspartate aminotransferase, and alkaline phosphatase) and cholesterol concentration increased significantly (p<0.05) in the group fed mold-contaminated feed while the serum albumin and total protein decreased significantly (p<0.05) in comparison with the negative control. Adverse histopathological changes were observed in the liver, kidney, spleen, and bursa of Fabricius in the group fed mold-contaminated feed. Addition of S. cerevisiae, bentonite or kaolin in the mold-contaminated feed ameliorated these toxic effects.

Conclusion: The observed histopathological lesions were consistent with mycotoxicosis in birds and were mild in the adsorbent treated groups. Kaolin had a higher protective effect against mycotoxicosis than the two other adsorbents.

Keywords: adsorbent, feed, mold, mycotoxin, pathology, Saccharomyces cerevisiae.

Stimulation of follicle growth and development during estrus in Ettawa Grade does fed a flushing supplement of different polyunsaturated fatty acids

Research (Published online: 05-01-2021)

2. Stimulation of follicle growth and development during estrus in Ettawa Grade does fed a flushing supplement of different polyunsaturated fatty acids

Prasetyo Nugroho, Komang Gede Wiryawan, Dewi Apri Astuti and Wasmen Manalu

Veterinary World, 14(1): 11-22

ABSTRACT

Background and Aim: Flushing with the manipulation of fatty acids, particularly polyunsaturated fatty acids, like linoleic and α-linolenic acids in the ration, is a strategy to raise the nutritional status of the female mammals to improve ovarian function and follicle development. This study was designed to investigate the effectiveness of flushing supplementation with different types of polyunsaturated and saturated fatty acids in stimulating follicle growth and development during estrus in Ettawa Grade does with a low initial body condition score (BCS ≤2).

Materials and Methods: Eighteen Ettawa Grade does in the second parity, with an average body weight of 32.11±2.19 kg, were divided into three groups according to the fatty acid supplemented to their ration: (i) About 2.8% lauric acid flushing (group); (ii) 2.8% linoleic acid flushing (LA group); and (iii) 2.8% a-linolenic acid flushing (ALA group). The ration was formulated to be isocaloric (total digestible nutrient = 77%) and isonitrogenous (crude protein = 15%). The experiment was conducted for 35 days; that is, 14 days for acclimatization and synchronization of the estrous cycle and 21 days for fatty acid flushing until the appearance of the next estrus. A completely randomized design was applied.

Results: According to the results, none of the different fatty acids in the ration affected the nutrient intakes, BCSs, average daily gains, and plasma glucose, cholesterol, and progesterone concentrations of the three groups of does. However, the BCSs (by 0.8-0.9) and the plasma cholesterol concentrations were higher after fatty acid flushing for 21 days than before the flushing period. The ALA group had the highest number of large-sized preovulatory follicles, whereas the LAURIC group had the highest plasma estradiol concentration during estrus. All three groups had similar plasma progesterone concentrations during estrus after fatty acid flushing.

Conclusion: Flushing supplementation with 2.8% ALA from flaxseed oil gave the best results in terms of stimulating the highest number of large-sized preovulatory follicles in Ettawa Grade does.

Keywords: Ettawa Grade does, flushing, follicle, lauric acid, linoleic acid, α-linolenic acid.

Seroprevalence and risk factors of brucellosis and the brucellosis model at the individual level of dairy cattle in the West Bandung District, Indonesia

Research (Published online: 05-01-2021)

1. Seroprevalence and risk factors of brucellosis and the brucellosis model at the individual level of dairy cattle in the West Bandung District, Indonesia

Yuli Yanti, Bambang Sumiarto, Tri Anggraeni Kusumastuti, Aprizal Panus and Sodirun Sodirun

Veterinary World, 14(1): 1-10

ABSTRACT

Background and Aim: Brucellosis is a zoonotic bacterial infectious disease. West Bandung is a center for dairy farming in West Java Province District and endemic for brucellosis. The aim of the study was to determine the prevalence, the associated risk factors, and the brucellosis model at the individual level of dairy cattle in the West Bandung District.

Materials and Methods: The research was conducted through a cross-sectional study. The samples were collected from the serum blood of dairy cattle. Data obtained from the questionnaire were used to investigate risk factors. Multistage random sampling was applied as the sampling technique; therefore, a sample size of 540 cows was selected. The number of farms and cattle on each farm was calculated using a variant effect design of the farm as well as 108 farms was selected with five cattle samples per farm. The results in regard of the research sample acquisition in the West Bandung District included 588 dairy cows from 116 farms, exceeds the number of samples calculated (540 dairy cows and 108 farms). The rose Bengal test (RBT) and the complement fixation test (CFT) were performed for brucellosis testing. Data associated with brucellosis cases at the individual level of the dairy cattle were analyzed using descriptive statistics univariate, bivariate with Chi-square, and odds ratio (OR). Moreover, multivariate logistic regression was used for the analysis during modeling.

Results: The results showed that the prevalence of brucellosis at the individual level in the West Bandung District was 5.10%. Risk factors associated with brucellosis in cattle included the history of abortion (p=0.000; OR=9.9), the history of placental retention (p=0.000; OR=6.6), the history of endometritis (p=0.000; OR=5.5), the history of stillbirth (p=0.043; OR=3.0), the history of pregnancy abortion age at 7-8 months (p=0.000; OR=15.2), and the history of pregnancy abortion at the age of 4-6 months (p=0.007; OR=3.8). The model of brucellosis in dairy cows was the following: = –3.2843+3.41033 the history of pregnancy abortion at the age of 7-8 months +2.54503 the history of pregnancy abortion at the age of 4-6 months +1.86185 age of cattle >2 years – 1.0469 Calving interval 12 months. The model showed the factors that were associated with brucellosis at the individual level of dairy cattle included the history of pregnancy abortion at the age of 7-8 months (β=+3.41033; OR=30.3), the history of pregnancy abortion at the age of 4-6 months (β=+2.54503; OR=12.7); age of cattle >2 years (β=+1.86185; OR=1.2), and Calving interval ≤12 months (β=–1.04691; OR=0.34).

Conclusion: The results of this research showed that the prevalence of brucellosis at the individual level of dairy cattle in the West Bandung district was 5.10%. The risk factors could contribute to the increase of the brucellosis cases, that is, the history of pregnancy abortion at the age of 7-8, the history of pregnancy abortion at the age of 4-6 months, and the age of cattle >2 years. The risk factors can be decreased in the brucellosis cases, that is, calving intervals ≤12 months.

Keywords: brucellosis, modeling, prevalence, risk factors.