RESUMO
On the basis of comparisons between bovine and ovine genome mapping information, the aim of the study was to analyze the genetic diversity of selected DNA microsatellites from the bovine genome and to investigate their correlation with the average daily milk yield in Awassi sheep. 18 informative microsatellite markers were selected from the significant QTL regions affecting milk yield identified in the bovine genome in previous studies. The selected microsatellite markers were then amplified by PCR as reciprocal amplifications on the genomic DNA of Awassi sheep, with standard daily milk yield records. Thus, in this study, 18 microsatellite markers associated with milk yield in the bovine genome were examined for both determination of genetic polymorphism within the flock and the effects of marker loci on average daily milk yield in Awassi sheep. Allele frequencies of markers were determined based on the results of fragment analysis. The analysis of variance showed that the 123 bp allele at the marker locus BMS1341 on BTA2 significantly influenced the average daily milk yield of Ivesi sheep (P < 0.01). On the other hand, the BMS381 locus with a 115 bp allele on BTA2, the MCM140 locus with a 185 bp allele on BTA6, the BMS2721 locus with a 155 bp allele, the BM1237 locus with 174 and 180 bp alleles on BTA7, and finally, the BMS1967 locus with a 117 bp allele, the BM4208 locus with 176 and 182 bp alleles, and the INRA locus with a185 bp allele on BTA8 showed moderately significant effects on the average daily milk yield of Ivesi ewes (P < 0.05).
Assuntos
Repetições de Microssatélites , Leite , Animais , Feminino , Turquia , Leite/metabolismo , Leite/química , Carneiro Doméstico/genética , Carneiro Doméstico/fisiologia , Locos de Características Quantitativas , Lactação , Frequência do Gene , Polimorfismo Genético , Reação em Cadeia da Polimerase/veterinária , Ovinos/genética , Bovinos/genética , Bovinos/fisiologiaRESUMO
Diarrhea caused by zoonotic pathogens is one of the most common diseases in dairy calves, threatening the health of young animals. Humans are also at risk, in particular children. To explore the pathogens causing diarrhea in dairy calves, the present study applied PCR-based sequencing tools to investigate the occurrence and molecular characteristics of three parasites (Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi) and three bacterial pathogens (Escherichia coli, Clostridium perfringens, and Salmonella spp.) in 343 fecal samples of diarrheic dairy calves from five farms in Lingwu County, Ningxia Hui Autonomous Region, China. The total positive rate of these pathogens in diarrheic dairy calves was 91.0% (312/343; 95% CI, 87.9-94.0), with C. perfringens (61.5%, 211/343; 95% CI, 56.3-66.7) being the dominant one. Co-infection with two to five pathogens was found in 67.3% (231/343; 95% CI, 62.4-72.3) of investigated samples. There were significant differences (p < 0.05) in the positive rates of Cryptosporidium spp. and diarrheagenic E. coli among farms, age groups, and seasons. Two Cryptosporidium species (C. parvum and C. bovis) and five gp60 subtypes of C. parvum (IIdA15G1, IIdA20G1, IIdA19G1, IIdA14G1, and a novel IIdA13G1) were identified. Two assemblages (assemblage E and zoonotic assemblage A) of G. duodenalis and six ITS genotypes of E. bieneusi (J, Henan-IV, EbpC, I, EbpA, and ESH-01) were observed. Four virulence genes (eaeA, stx1, stx2, and st) of diarrheagenic E. coli and one toxin type (type A) of C. perfringens were detected. Our study enriches our knowledge on the characteristics and zoonotic potential of diarrhea-related pathogens in dairy calves.
Title: Caractérisation moléculaire des protozoaires parasites zoonotiques courants et des bactéries responsables de diarrhée chez les veaux laitiers dans la région autonome Hui du Ningxia, en Chine. Abstract: La diarrhée causée par des agents pathogènes zoonotiques est l'une des maladies les plus courantes chez les veaux laitiers, menaçant la santé des jeunes animaux. Ceci est également un risque pour la santé humaine, en particulier les enfants. Pour explorer les agents pathogènes responsables de la diarrhée chez les veaux laitiers, cette étude a utilisé des outils de séquençage basés sur la PCR pour étudier l'occurrence et les caractères moléculaires de trois parasites (Cryptosporidium spp., Giardia duodenalis et Enterocytozoon bieneusi) et de trois agents pathogènes bactériens (Escherichia coli, Clostridium perfringens et Salmonella spp.) dans 343 échantillons fécaux de veaux laitiers diarrhéiques provenant de cinq fermes du comté de Lingwu, région autonome Hui du Ningxia, en Chine. Le taux total positif de ces pathogènes chez les veaux laitiers diarrhéiques était de 91,0 % (312/343; IC à 95 %, 87,994,0), et C. perfringens (61,5 %, 211/343; IC à 95 %, 56,366,7) était le plus répandu. Une co-infection avec deux à cinq pathogènes a été trouvée dans 67,3 % (231/343; IC à 95 %, 62,472,3) des échantillons étudiés. Il y avait des différences significatives (p < 0,05) dans les taux positifs de Cryptosporidium spp. et d'E. coli diarrhéogènes entre les fermes, les groupes d'âge et les saisons. Deux espèces de Cryptosporidium (C. parvum et C. bovis) et cinq sous-types de gp60 de C. parvum (IIdA15G1, IIdA20G1, IIdA19G1, IIdA14G1 et un nouveau, IIdA13G1) ont été identifiés. Deux assemblages (assemblage E et assemblage zoonotique A) de G. duodenalis et six génotypes ITS d'E. bieneusi (J, Henan-IV, EbpC, I, EbpA et ESH-01) ont été observés. Quatre gènes de virulence (eaeA, stx1, stx2 et st) d'E. coli diarrhéogènes et un type de toxine (type A) de C. perfringens ont été détectés. Notre étude enrichit les connaissances sur les caractères et le potentiel zoonotique des agents pathogènes liés à la diarrhée chez les veaux laitiers.
Assuntos
Doenças dos Bovinos , Criptosporidiose , Cryptosporidium , Diarreia , Enterocytozoon , Fezes , Giardia lamblia , Zoonoses , Animais , Bovinos , Diarreia/veterinária , Diarreia/parasitologia , Diarreia/microbiologia , Diarreia/epidemiologia , China/epidemiologia , Doenças dos Bovinos/parasitologia , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/microbiologia , Cryptosporidium/genética , Cryptosporidium/isolamento & purificação , Cryptosporidium/classificação , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , Enterocytozoon/classificação , Giardia lamblia/genética , Giardia lamblia/isolamento & purificação , Giardia lamblia/classificação , Fezes/parasitologia , Fezes/microbiologia , Criptosporidiose/epidemiologia , Criptosporidiose/parasitologia , Escherichia coli/isolamento & purificação , Escherichia coli/genética , Escherichia coli/classificação , Giardíase/veterinária , Giardíase/epidemiologia , Giardíase/parasitologia , Coinfecção/veterinária , Coinfecção/epidemiologia , Coinfecção/parasitologia , Coinfecção/microbiologia , Microsporidiose/veterinária , Microsporidiose/epidemiologia , Clostridium perfringens/isolamento & purificação , Clostridium perfringens/genética , Clostridium perfringens/classificação , Salmonella/isolamento & purificação , Salmonella/genética , Salmonella/classificação , Humanos , Reação em Cadeia da Polimerase/veterinária , Indústria de LaticíniosRESUMO
The bovine leukocyte antigen (BoLA) gene is a significant genetic part of the immune system and has been used as a disease marker in cattle. In this study, we detected Theileria orientalis, T. sinensis, Anaplasma marginale, Anaplasma platys, Candidatus Mycoplasma haemobos and Trypanosoma evansi by PCR amplification and sequencing of the amplicons. The allelic association of the BoLA-DRB3.2 gene with blood pathogen disease resistance and susceptibility in 87 Kedah-Kelantan x Brahman (KKB) and 38 Bali cattle was determined by Fisher's exact test and Cochran Mantel Haenszel (CMH) correction test. Sequence-based typing of the BoLA-DRB3.2 gene identified 43 alleles (27 previously reported alleles and 16 novel alleles) across the two cattle breeds. Alignment analysis of the 16 novel alleles revealed 90.7-95.8% and 85-92% nucleotide and amino acid identities, with the reference allele, BoLA-DRB3*016:01 cDNA clone NR-1. BoLA-DRB3*009:02 (25.6%) and BoLA-DRB3*036:01 (36%) were the most frequent alleles in KKB and Bali cattle, respectively. In KKB cattle, BoLA-DRB3*020:02:01 was significantly associated with resistance to T. orientalis whereas *007:01 and *009:02 were significantly associated with resistance to C. Mycoplasma haemobos. Also, DRB3*017:01 was associated with susceptibility to T. orientalis in KKB cattle. In the Bali cattle, BoLA-DRB3*015:01 was found to be a genetic marker of susceptibility to C. Mycoplasma haemobos infection. Therefore, this study identified BoLA-DRB3.2 alleles associated with resistance and susceptibility to T. orientalis infection in KKB cattle and susceptibility to C. Mycoplasma haemobos infection in Bali cattle for the first time. Therefore, this study suggests that these BoLA-DRB3 resistance alleles could be used as candidate markers for selection, whereas susceptibility alleles could be used as candidate markers for culling in the beef industry.
Assuntos
Alelos , Resistência à Doença , Theileria , Theileriose , Animais , Bovinos , Theileriose/parasitologia , Theileria/genética , Projetos Piloto , Resistência à Doença/genética , Antígenos de Histocompatibilidade Classe II/genética , Doenças dos Bovinos/parasitologia , Doenças dos Bovinos/genética , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/imunologia , Índia , Reação em Cadeia da Polimerase/veterináriaRESUMO
Vector-borne pathogens continue to increase their impact on the livestock industry worldwide. To protect animals against these pathogens, it is very important to identify the species that cause the disease and understand their prevalence. This study aimed to investigate the presence and prevalence of vector-borne pathogens in apparently healthy cattle in different parts of Kyrgyzstan using molecular diagnostic techniques. For this purpose, 531 blood samples were collected from the Osh, Jalal-Abad, and Batken oblasts of Kyrgyzstan. The blood samples were investigated for vector-borne pathogens using PCR, RLB, and RFLP. Moreover, DNA sequence analyses were used to confirm the results of molecular techniques and phylogenetic analyses of these pathogens. 359 (67.61%) out of 531 samples were found to be infected with at least one pathogen, whereas 172 (32.39%) were detected to be negative. Thirteen vector-borne pathogens were detected in cattle blood samples, and the prevalence of these pathogens was as follows: Theileria orientalis (47.83%), T. annulata (25.61%), Babesia major (0.19%), B. occultans (0.38%), Anaplasma phagocytophilum-like 1 (3.20%), A. capra (3.01%), A. centrale (2.82%), A. bovis (1.13%), (A) ovis (0.19%), Candidatus Anaplasma camelii (0.94%), Trypanosoma theileri (19.21%), Mycoplasma wenyonii (6.03%), and Ca. Mycoplasma haemobos (2.64%). Among the positive samples, one pathogen was identified in 189 cattle (35.59%), and co-infections (two or more pathogens) were determined in 170 (32.01%) animals. Theileria parva, T. mutans, (B) bigemina, B. bovis, B. divergens, and A. marginale could not be detected in the study. Anaplasma bovis and Ca. Anaplasma camelii were detected for the first time in the country. This molecular survey provides important epidemiological and genetic data for the vector-borne pathogens in cattle. The results of the study showed that vector-borne pathogens have a significant spread and distribution in cattle in Kyrgyzstan.
Assuntos
Anaplasma , Anaplasmose , Doenças dos Bovinos , Animais , Bovinos , Quirguistão/epidemiologia , Anaplasma/isolamento & purificação , Anaplasma/genética , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/parasitologia , Anaplasmose/epidemiologia , Anaplasmose/microbiologia , Prevalência , Filogenia , Reação em Cadeia da Polimerase/veterinária , Theileria/isolamento & purificação , Theileria/genética , Theileriose/epidemiologia , Theileriose/parasitologia , Theileriose/sangue , Polimorfismo de Fragmento de RestriçãoRESUMO
Background: Milk and its products are very sensitive to spoilage if they are kept under unsuitable conditions which may provide favorable circumstances for the growth of specific spoilage organisms, Pseudomonas fluorescens accounted as the most dominant indicator for milk spoilage. Aim: This study highlights monitoring the prevalence of P. fluorescens as a spoilage indicator organism in cow raw milk and its contact surfaces represented by teat surfaces and milk tanks in Nineveh province. Methods: A total of 150 samples from cows' raw milk, teat surfaces, and milk tank swabs were collected from different locations in Nineveh province from October 2023 till February 2024. The Pseudomonas fluorescens were detected by using conventional cultivation methods supported by molecular detection of the target pathogen using the polymerase chain reaction technique. Results: Out of 150 samples, 48 (32%) were positive for the prevalence of P. fluorescens by traditional methods, and 39 (26%) were positive using PCR assay according to the 16SPflu gene yielded a band at 850 bp. The P. fluorescens was recovered at 19 (38%) from raw milk. Teat surfaces revealed a higher isolation rate 11 (22%) compared to milk tanks 9 (18%). The mean counts of Pseudomonas in cows raw milk revealed 4.38, 6.29, and 7.37 log CFU/ml for the 0, 3, and 6 days of storage at chilling temperature. Results of DNA sequencing of the 16SrRNA gene revealed 12 strains recorded in the GenBank nucleotide sequence database. Conclusion: Our results shed light on the risk of P. fluorescens prevalence as a spoilage indicator in raw milk and surrounding surfaces which is inevitable to apply hygienic procedures during milk collecting, processing, and preservation to increase the shelf life of the products and ensure milk safety and consumer health.
Assuntos
Leite , Pseudomonas fluorescens , Animais , Pseudomonas fluorescens/isolamento & purificação , Leite/microbiologia , Bovinos , Feminino , Glândulas Mamárias Animais/microbiologia , Prevalência , Microbiologia de Alimentos , Reação em Cadeia da Polimerase/veterináriaRESUMO
Background: Dairy products are considered some important sources of various nutritional compounds; however, pathogenic bacterial growth is a critical destructive factor to these products leading to consumer health and system financial crises. Aim: The current study was carried out to identify if there is any presence of Staphylococcus aureus-related enterotoxin genes in cheese samples. Methods: The research included the collection of 35 samples. The samples passed through conventional cultivation processes and a PCR method to detect the presence of icaA, sea, hla, and fnbA enterotoxin genes in these samples. Results: The conventional identification revealed the growth of S. aureus from the cheese samples. The PCR findings recorded the presence of the icaA, sea, hla, and fnbA in 31 (88.5%), 27 (77%), 19 (54%), and 12 (34%), respectively, of cheese samples. The sequencing revealed close similarities with global isolates, which reached up to 98.5% of identity. Conclusion: The current results indicate the presence of enterotoxin genes of S. aureus in high rates in the dairy products examined, which reveals critical problems of food safety due to the possible presence of enterotoxins in consumer dairy products.
Assuntos
Queijo , Enterotoxinas , Staphylococcus aureus , Queijo/microbiologia , Enterotoxinas/genética , Staphylococcus aureus/genética , Staphylococcus aureus/isolamento & purificação , Microbiologia de Alimentos , Reação em Cadeia da Polimerase/veterináriaRESUMO
Background: Lumpy skin disease (LSD) is caused by a virus belonging to the genus Capripoxvirus, exhibiting clinical symptoms ranging from mild signs to the development of nodules. LSD emerged in Asia and Southeast Asia, including Vietnam, in October 2020 and has since spread throughout the region, resulting in productivity and economic losses. Aim: This study aimed to investigate the virus-causing papular dermatitis in cattle from the Mekong Delta region of Vietnam by analyzing its GPCR gene and assessing its evolutionary relationship with sequences in the GenBank database. Methods: Blood samples (n = 180) were collected from cattle farms in Ben Tre, Tien Giang, and Tra Vinh provinces. PCR targeting the P32 antigen gene was utilized to detect LSDV presence, and GPCR gene amplification was performed to assess genetic variability. Results: LSDV was detected in 8.33% (15/180) of the samples using PCR targeting the P32 antigen gene. Each sample that tested positive for LSDV demonstrated complete amplification of the GPCR gene. Sequence alignments and phylogenetic analyses of the GPCR gene revealed that Mekong Delta LSDV isolates shared genetic similarities and possessed a 12-nucleotide insertion comparable to strains from China in 2019 and Northern Vietnam in 2020. Conclusion: This study provides preliminary insights into the molecular characteristics of LSDV in cattle from the Mekong Delta region of Vietnam. The observed genetic relatedness to other LSDV sequences from Asia and Southeast Asia underscores the importance of regional surveillance and control measures. These findings contribute to the development of effective strategies for LSDV control and prevention.
Assuntos
Doença Nodular Cutânea , Vírus da Doença Nodular Cutânea , Filogenia , Animais , Bovinos , Vietnã/epidemiologia , Doença Nodular Cutânea/virologia , Doença Nodular Cutânea/epidemiologia , Vírus da Doença Nodular Cutânea/genética , Vírus da Doença Nodular Cutânea/isolamento & purificação , Reação em Cadeia da Polimerase/veterináriaRESUMO
Background: Tick is one of the most important ectoparasites distributed worldwide and plays an obvious role in the transmission of different infections to humans and animals as dogs. Aim: This study conducted to molecular demonstration of Babesia gibsoni in ticks of stray dogs and phylogenetic analysis of study isolates to detect their identity to global isolates. Prevalence of ticks in dogs, identification of tick species, and their relationship to some risk factors were aimed, also. Methods: A total of 97 stray dogs were inspected grossly to detect and collect ticks that existed in different body parts. After collection, all ticks were examined morphologically to identify their species, and then molecularly by the polymerase chain reaction (PCR) assay to detect B. gibsoni in different species of ticks. Local B. gibsoni isolates were sequenced, documented in the National Center For biotechnology information (NCBI) database, analyzed phylogenetically, and compared with the global GenBank-NCBI isolates. Results: In the current study, ticks were detected in 43.3% of dogs, and were shown to be varied in number and distribution among different body parts of each dog. Concerning its distribution, ticks were observed significantly on the abdomen, ear, and perineal region. In relation to risk factors, ticks were increased significantly in dogs <6 months old in comparison to older dogs, males more than females; and in rural areas more than dogs of sub-urban and urban areas. Based on morphology, different tick species were seen including Hylaomma anatolicum (86.12%), R. sanguineus (11.99%), and Rhipicephalus turanicus (1.89%). Targeting the 18S rRNA gene, PCR assay reported 3.79% positive ticks to B. gibsoni that were seen in R. sanguineus (13.16%) and H. anatolicum (2.56%). Based on phylogenetic analysis data of five local B. gibsoni isolates, this study demonstrated their close relations to the global NCBI-BLAST B. gibsoni Iraqi isolate (ID: MN385424.1). Conclusion: This represents the first Iraqi study that demonstrated molecularly B. gibsoni in different species of ticks that infected stray dogs.
Assuntos
Babesia , Babesiose , Doenças do Cão , Filogenia , Carrapatos , Animais , Cães , Babesia/isolamento & purificação , Babesia/genética , Doenças do Cão/parasitologia , Doenças do Cão/epidemiologia , Iraque/epidemiologia , Masculino , Babesiose/epidemiologia , Babesiose/parasitologia , Feminino , Carrapatos/parasitologia , Infestações por Carrapato/veterinária , Infestações por Carrapato/epidemiologia , Infestações por Carrapato/parasitologia , Prevalência , Reação em Cadeia da Polimerase/veterináriaRESUMO
Herein we describe a single nucleotide polymorphism-specific polymerase chain reaction (PCR) assay to rapidly detect and differentiate variants belonging to the European and North American lineages of Echinococcus multilocularis in clinical samples. This is an extremely relevant and applicable test in North America because the range of E. multilocularis continues to expand across the continent and because of a rise in prevalence in wildlife, domestic animals, and humans. The endemic North American (NA) and introduced European (EU) variants are believed to have different pathogenic potentials, with the EU variants being more infective and pathogenic than the NA variants. The rise of the EU variants of E. multilocularis increases the risk of spillover from wildlife to humans because of its increased potential for infectivity. Current PCR-based diagnostics can detect E. multilocularis deoxyribonucleic acid (DNA), but DNA sequencing is required to identify the specific variant. Our assay provides a straightforward conventional PCR method to differentiate the NA and EU variants, and we suggest this same approach could be used for the diagnosis of other parasites or variants that are genetically very similar. As surveillance continues for E. multilocularis across North America, identifying the different genetic variants from different geographic regions will become essential to understanding the current epidemiological shift that the parasite is experiencing, as well as informing public health decisions in affected areas.
Assuntos
DNA de Helmintos , Equinococose , Echinococcus multilocularis , Haplótipos , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único , Echinococcus multilocularis/genética , Echinococcus multilocularis/classificação , Echinococcus multilocularis/isolamento & purificação , Animais , Reação em Cadeia da Polimerase/veterinária , Reação em Cadeia da Polimerase/métodos , Equinococose/parasitologia , Equinococose/veterinária , Equinococose/diagnóstico , Equinococose/epidemiologia , Europa (Continente)/epidemiologia , América do Norte/epidemiologia , HumanosRESUMO
PURPOSE: Theileriosis in cattle has a significant economic implication for dairy production globally. Thus, it is crucial to investigate the prevalence of bovine theileriosis, the causative agent and genotypes of Theileria species in dairy cattle in the Kurunegala District in the Intermediate zone, where the third largest population of dairy cattle in Sri Lanka is present and experienced a substantial reduction in the cattle population recently. METHODS: Sixty blood samples and background data were collected from three dairy farms in Galpokuna (n = 20), Koulwewa (n = 20), and Andigama (n = 20) areas. Haematocrit was used to identify anaemia while Giemsa-stained blood smears demonstrated the presence of piroplasms. A Fisher's Exact Test (p < 0.05) compared the prevalence of infection among age groups and farms. PCRs using species-specific primers designed to amplify regions of MPSP and 18 s rRNA genes followed by sequencing of selected amplicons allowed phylogeny of the species detected. RESULTS: All three farms had semi-intensive farming practices keeping animals in a 'closed' setup with limited movement. Theileriosis prevalence in dairy cattle was 55.0 % with no difference among the farms (Koulwewa: 65.0 %, Galpokuna: 50.0 %, Andigama: 50.0 %). One-third of the cattle (33.3 %) were anaemic based on haematocrit results but most showed mild anaemia. Anaemia was significantly higher in calves (45.0 %) than in adults (25.0 %; χ2 = 5.743; p = 0.03) tested positive for theileriosis. None of the animals showed any other signs of theileriosis. PCR revealed the presence of both T. orientalis (pathogenic or benign) and T. annulata (pathogenic). The sequencing data revealed that the T. orientalis genotype present in Kurunegala District is type 7. CONCLUSION: This is the first report on T. orientalis and T. annulata in dairy cattle in the Intermediate zone of Sri Lanka. The type 7 of T. orientalis was more common showing severe anaemia in calves while mild anaemia or no anaemia in adults and sub-adults. In immunologically intact animals, the reported genotype of the parasite can persist asymptomatically for life, occasionally causing a relapse, particularly under stressful conditions like pregnancy, lactation, and rapid changes in environmental conditions. However, the susceptibility of calves for pathogenic and apathogenic genotypes of T. orientalis and the carrier status of asymptomatic animals needs further investigation.
Assuntos
Genótipo , Theileria , Theileriose , Animais , Theileriose/epidemiologia , Theileriose/parasitologia , Bovinos , Sri Lanka/epidemiologia , Theileria/isolamento & purificação , Theileria/genética , Theileria/classificação , Prevalência , Feminino , Filogenia , Indústria de Laticínios , Reação em Cadeia da Polimerase/veterinária , Anemia/veterinária , Anemia/parasitologia , Anemia/epidemiologiaRESUMO
This study examined Haemonchus contortus prevalence and benzimidazole resistance in eight districts of Marathwada, Maharashtra, India. A comprehensive investigation of 264 abomasa of goats collected from abattoirs and goats necropsied at the College of Veterinary Sciences and Animal Husbandry, Parbhani, revealed 21.21 % a prevalence of H. contortus. The incidence of H. contortus did not vary much across seasons and it was highest in summer (23.42 %), followed by monsoon (22.89 %), and lowest in winter (15.71 %). Statistically non-significant (p < 0.05) prevalence was observed in male and female animals. A detailed examination of 168 adult H. contortus worms from eight districts revealed the presence of all conceivable genotypes including homozygous resistant (rr), susceptible (SS), and heterozygous (Sr) BZ susceptible genotypes. The rr was the most frequent at 50 %, followed by SS at 27 % and Sr at 22 %. The presence of the SNP was observed in in all eight randomly selected and sequenced samples.
Assuntos
Benzimidazóis , Resistência a Medicamentos , Doenças das Cabras , Cabras , Hemoncose , Haemonchus , Animais , Haemonchus/efeitos dos fármacos , Haemonchus/genética , Doenças das Cabras/parasitologia , Doenças das Cabras/epidemiologia , Índia/epidemiologia , Benzimidazóis/farmacologia , Hemoncose/veterinária , Hemoncose/parasitologia , Hemoncose/epidemiologia , Prevalência , Feminino , Masculino , Genótipo , Reação em Cadeia da Polimerase/veterinária , Anti-Helmínticos/farmacologia , Anti-Helmínticos/uso terapêutico , Estações do AnoRESUMO
Babesia bigemina and Theileria annulata are tick-borne protozoans that cause piroplasmosis in cattle, resulting in huge damages to the livestock industry. The prevalence of these infections depends on various intrinsic and extrinsic risk factors. In Pakistan, there is no information regarding the molecular characterization of Babesia bigemina and the risk factors associated with piroplasmosis. This study aimed to molecularly characterize Babesia spp. and Theileria spp. infecting various cattle breeds in Khyber Pakhtunkhwa, Pakistan, and to shed light on risk factors associated with these infections. Altogether, 219 blood samples were collected from various symptomatic cattle breeds, including Holstein Friesian (65.3%; 143/219), Jersey (21.5%; 47/219) and Sahiwal (13.2%; 29/219). Isolated genomic DNA from these blood samples was used in PCR for the amplification of the 18S rRNA fragment of apicomplexan parasites. Obtained 18S rDNA sequences from cattle hosts showed 99.5% identity with B. bigemina, or 100% with T. annulata. Having an overall infection rate of 61.6% (135/219), the highest infection rate was recorded for T. annulata (43.8%; 95/219), followed by B. bigemina (18.3%; 40/219). Phylogenetic analysis of 18S rDNA sequences revealed that B. bigemina clustered with corresponding species reported from Bolivia, and South Africa, while T. annulata grouped with same species from Italy, India, and Turkey. Among the different risk factors, the breed, season, and tick infestation were found to have a significant (P < 0.05) association with the piroplasmid infections. The information obtained in this study can be employed for effective surveillance and control of babesiosis and theileriosis in Pakistan. In addition to confirming our previous molecular detection of T. annulata in cattle, this study provides the first molecular surveillance and phylogenetic position of B. bigemina and associated risk factors in the study region.
Assuntos
Babesia , Babesiose , Doenças dos Bovinos , Filogenia , RNA Ribossômico 18S , Theileria annulata , Theileriose , Bovinos , Animais , Babesia/isolamento & purificação , Babesia/genética , Babesia/classificação , Theileriose/epidemiologia , Theileriose/parasitologia , Babesiose/epidemiologia , Babesiose/parasitologia , Theileria annulata/genética , Theileria annulata/isolamento & purificação , Fatores de Risco , Paquistão/epidemiologia , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/parasitologia , RNA Ribossômico 18S/análise , RNA Ribossômico 18S/genética , Prevalência , DNA de Protozoário/análise , Reação em Cadeia da Polimerase/veterinária , FemininoRESUMO
BACKGROUND: Due to the diversity of Shiga toxin-producing Escherichia coli (STEC) isolates, detecting highly pathogenic strains in foodstuffs is challenging. Currently, reference protocols for STEC rely on the molecular detection of eae and the stx1 and/or stx2 genes, followed by the detection of serogroup-specific wzx or wzy genes related to the top 7 serogroups. However, these screening methods do not distinguish between samples in which a STEC possessing both determinants are present and those containing two or more organisms, each containing one of these genes. This study aimed to evaluate ecf1, Z2098, Z2099, and nleA genes as single markers and their combinations (ecf1/Z2098, ecf1/Z2099, ecf1/nleA, Z2098/Z2099, Z2098/nleA, and Z2099/nleA) as genetic markers to detect potentially pathogenic STEC by the polymerase chain reaction (PCR) in 96 animal samples, as well as in 52 whole genome sequences of human samples via in silico PCR analyses. RESULTS: In animal isolates, Z2098 and Z2098/Z2099 showed a strong association with the detected top 7 isolates, with 100% and 69.2% of them testing positive, respectively. In human isolates, Z2099 was detected in 95% of the top 7 HUS isolates, while Z2098/Z2099 and ecf1/Z2099 were detected in 87.5% of the top 7 HUS isolates. CONCLUSIONS: Overall, using a single gene marker, Z2098, Z2099, and ecf1 are sensitive targets for screening the top 7 STEC isolates, and the combination of Z2098/Z2099 offers a more targeted initial screening method to detect the top 7 STEC isolates. Detecting non-top 7 STEC in both animal and human samples proved challenging due to inconsistent characteristics associated with the genetic markers studied.
Assuntos
Escherichia coli Êntero-Hemorrágica , Infecções por Escherichia coli , Escherichia coli Shiga Toxigênica , Escherichia coli Shiga Toxigênica/genética , Escherichia coli Shiga Toxigênica/isolamento & purificação , Animais , Marcadores Genéticos , Infecções por Escherichia coli/veterinária , Infecções por Escherichia coli/microbiologia , Escherichia coli Êntero-Hemorrágica/genética , Escherichia coli Êntero-Hemorrágica/isolamento & purificação , Humanos , Plasmídeos/genética , Simulação por Computador , Bovinos , Reação em Cadeia da Polimerase/veterinária , Ovinos , Ilhas Genômicas/genéticaRESUMO
Toxoplasmosis is one of the most common parasitic zoonoses and represents a significant health risk for humans, especially for immunodeficient patients. The main transmission route is by oral uptake of oocysts and consumption of undercooked meat of infected animals. Different species have been evaluated as possible reservoirs of the parasite, but few studies have been carried out to examine the role of horses in transmission of the disease. Given the proximity of these animals to humans and the widespread consumption of their meat in many countries, including the Mediterranean basin, it is important to determine the prevalence of T. gondii infection in this species. In this study, blood samples from 105 horses were collected and the presence of T. gondii was evaluated by serological and molecular methods. Antibodies against T. gondii of 12 horses (11.43%) were detected by enzyme-linked immunosorbent assay (ELISA), whereas 29 horses (27.62%) showed positive for PCR. Seroprevalence was related to use of the animals, being higher in horses used for dressage than in others. Purebreds had higher seroprevalence than crossbred animals. No differences between breed, sex or age were found. The results of this study confirm the presence of T. gondii infection in horses, highlighting the need to analyse the meat of this species before human consumption and to control of this infection in horses, as they could be an important reservoir of this zoonotic parasite.
Assuntos
Anticorpos Antiprotozoários , Ensaio de Imunoadsorção Enzimática , Doenças dos Cavalos , Toxoplasma , Toxoplasmose Animal , Animais , Cavalos/parasitologia , Toxoplasmose Animal/epidemiologia , Toxoplasmose Animal/diagnóstico , Toxoplasmose Animal/parasitologia , Espanha/epidemiologia , Doenças dos Cavalos/parasitologia , Doenças dos Cavalos/epidemiologia , Doenças dos Cavalos/diagnóstico , Toxoplasma/isolamento & purificação , Toxoplasma/genética , Estudos Soroepidemiológicos , Feminino , Anticorpos Antiprotozoários/sangue , Masculino , Ensaio de Imunoadsorção Enzimática/veterinária , Reação em Cadeia da Polimerase/veterinária , PrevalênciaRESUMO
BACKGROUND: Hepatozoonosis has been reported in many species around the world. Few incidences have been reported in various species of wild felids. Tigers are endangered large cats and are protected under the Wildlife Protection Act, 1972 under Schedule I. The study was carried out to estimate the positivity rate of hepatozoonosis in tigers of the Vidarbha region of Maharashtra, India. METHODS: Blood (n = 21) or tissue samples (n = 5) were collected from 26 wild captured / zoo-born or dead tigers during the quarantine period/post-mortem examination. Blood smear examination along with Polymerase Chain Reaction (PCR) studies were conducted for the detection of hepatozoonosis. All the amplicons from the positive samples were purified and sequenced, and the sequences were subjected to nBLAST analysis to detect the species of Hepatozoon. The sequences were deposited into public domain database of National Center for Biotechnology Information (NCBI) and accession numbers were allotted. A phylogenetic study was undertaken to understand the evolutionary lineage of the pathogen. Tissue distribution studies were carried out on tissue samples received during post mortem. A clinical case in a tiger cub was managed and sub-clinical cases were monitored for relapse. Age-wise, sex-wise, region-wise and captive time-wise positivity rate was estimated. The data was analyzed using statistical tools. RESULTS: A total of 12 tigers were found positive for H. felis during the screening. A clinical case was diagnosed and successfully treated. The age group of 0-3 years reported a positivity rate of 66.66%, and all the cases found positive were reported between the age group of 0-7 years. Males reported a positivity rate of 58.33 per cent, while females reported 35.71%. Taboba and Andhari Tiger Reserve of the state had a positivity rate of 52.94 per cent. However, the statistical analysis for blood parameters and positivity rate by 't' test and Chi-squared test were found to be non-significant. CONCLUSIONS: An overall positivity rate of 46.15% indicates the wide distribution of hepatozoonosis among wild tigers of the Vidarbha region of Maharashtra, India, which is strategically important considering the gene flow and migration of tigers. Hepatozoonosis can progress to clinical outcomes in young animals and require veterinary intervention. Molecular tools and phylogenetic studies can supplement important data on circulating species of Hepatozoon in the field. Further studies on the clinical management and epidemiology of the infection in wild felids will comprehend the cause of wildlife conservation.
Assuntos
Coccidiose , Filogenia , Tigres , Animais , Índia/epidemiologia , Coccidiose/veterinária , Coccidiose/epidemiologia , Coccidiose/parasitologia , Tigres/parasitologia , Masculino , Feminino , Eucoccidiida/genética , Eucoccidiida/isolamento & purificação , Reação em Cadeia da Polimerase/veterináriaRESUMO
Feline leukemia virus (FeLV) is a highly debilitating cat pathogen due to its ability to cause many pathological changes. Therefore, identifying the virus directly in bone marrow can be a highly relevant diagnostic tool even in the absence of viraemia. The aim of this study was to compare the diagnostic efficiency of immunocytochemistry (ICC) of bone marrow aspirates with enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). Blood samples were collected from 188 cats and separated into aliquots of whole blood for nested PCR using the U3 LTR region and the gag gene of FeLV-A as reference and serum for detection of the p27 antigen by ELISA. Bone marrow samples from these cats were placed on silanized slides for anti-FeLV ICC using gp70 as primary antibody. A total of 28.2% of the cats tested for FeLV were positive in at least one of the tests, with 26.6% positive by PCR, 18.1% by ICC and 11.2% by ELISA. Cohen's kappa agreement test revealed moderate agreement between ELISA and PCR results and substantial agreement between ICC and ELISA and between ICC and PCR. The results indicated that ICC of bone marrow is an efficient novel diagnostic test for FeLV infection.
Assuntos
Medula Óssea , Ensaio de Imunoadsorção Enzimática , Imuno-Histoquímica , Vírus da Leucemia Felina , Gatos , Animais , Ensaio de Imunoadsorção Enzimática/veterinária , Medula Óssea/virologia , Leucemia Felina/diagnóstico , Infecções por Retroviridae/veterinária , Infecções por Retroviridae/diagnóstico , Reação em Cadeia da Polimerase/veterinária , Infecções Tumorais por Vírus/veterinária , Infecções Tumorais por Vírus/diagnóstico , Doenças do Gato/diagnóstico , Doenças do Gato/virologiaRESUMO
Malignant catarrhal fever (MCF) presents a sporadic yet significant threat to livestock and wildlife. A comprehensive investigation in Karnataka, India into the prevalence and transmission patterns of sheep-associated MCF (SA-MCF) was conducted. A total of 507 sheep peripheral blood leukocyte samples from 13 districts along with 27 cows and 10 buffalo samples from various regions in Karnataka were tested for SA-MCF infection i.e. Ovine gammaherpesvirus 2 (OvHV-2) using heminested PCR. Furthermore, serum samples collected from 73 cows and 15 buffalo suspected of MCF were tested using a commercially available ELISA kit. Additionally, histopathological examinations of affected tissues and phylogenetic analysis of viral tegument protein sequences were conducted. Our findings indicated a 20.11%, 33.33% and 20% positivity for OvHV-2 in sheep, cows and buffalo respectively by PCR. Statistical analysis revealed a significant association between the age of sheep and the detection of OvHV-2. Seven cows and one buffalo serum samples tested positive for ELISA. Clinical findings in bovids were consistent with typical MCF signs, and histopathological results revealed multi-organ involvement characterised by necrotising vasculitis and lymphoid hyperplasia. The nucleotide pairwise identity matrix revealed 99.5% identity between the sequences obtained in the study with sequences from other states. The phylogenetic analysis of partial tegument protein sequences from bovid and sheep samples suggested a close genetic relationship between the local OvHV-2 strains and those from various global regions. Crucially, this study underscores the widespread presence of SA-MCF in Karnataka, with significant implications for both livestock management and wildlife conservation.
Assuntos
Búfalos , Gammaherpesvirinae , Febre Catarral Maligna , Filogenia , Animais , Febre Catarral Maligna/virologia , Febre Catarral Maligna/transmissão , Febre Catarral Maligna/epidemiologia , Febre Catarral Maligna/patologia , Índia/epidemiologia , Ovinos , Gammaherpesvirinae/genética , Gammaherpesvirinae/isolamento & purificação , Gammaherpesvirinae/classificação , Bovinos , Búfalos/virologia , Doenças dos Ovinos/virologia , Doenças dos Ovinos/transmissão , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/patologia , Feminino , Prevalência , Ensaio de Imunoadsorção Enzimática/veterinária , Doenças dos Bovinos/virologia , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/transmissão , Doenças dos Bovinos/patologia , Reação em Cadeia da Polimerase/veterináriaRESUMO
Brucellosis represents a major public health concern worldwide. Human transmission is mainly due to the consumption of unpasteurized milk and dairy products of infected animals. The gold standard for the diagnosis of Brucella spp in ruminants is the bacterial isolation, but it is time-consuming. Polymerase Chain Reaction (PCR) is a quicker and more sensitive technique than bacterial culture. Droplet digital PCR (ddPCR) is a novel molecular assay showing high sensitivity in samples with low amount of DNA and lower susceptibility to amplification inhibitors. Present study aimed to develop a ddPCR protocol for the detection of Brucella abortus in buffalo tissue samples. The protocol was validated using proficiency test samples for Brucella spp by real time qPCR. Furthermore, 599 tissue samples were examined. Among reference materials, qPCR and ddPCR demonstrated same performance and were able to detect up to 225 CFU/mL. Among field samples, ddPCR showed higher sensitivity (100%), specificity and accuracy of 93.4% and 94.15%, respectively. ddPCR could be considered a promising technique to detect B. abortus in veterinary specimens, frequently characterized by low amount of bacteria, high diversity in matrices and species and poor storage conditions.
Assuntos
Brucella abortus , Brucelose , Búfalos , DNA Bacteriano , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e Especificidade , Animais , Brucella abortus/isolamento & purificação , Brucella abortus/genética , Búfalos/microbiologia , Brucelose/veterinária , Brucelose/diagnóstico , Brucelose/microbiologia , DNA Bacteriano/análise , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase/veterinária , Reação em Cadeia da Polimerase/métodosRESUMO
Mycoplasma bovis (M. bovis) is capable of causing a range of diseases in cattle, encompassing calf pneumonia, arthritis, conjunctivitis, meningitis, and mastitis. It is widely recognized as one of the predominant pathogens posing a significant threat to the global cattle industry. Therefore, accurate and sensitive methods are urgently needed to detect M. bovis. This study aims to detect M. bovis by combining colloidal gold with biotin-labeled oligonucleotides to improve detection sensitivity and form a chromogenic detection probe based on signal amplification technology. Here, we developed a sensitive and specific polymerase chain reaction-lateral flow dipstick assay (PCR-LFD) strip for efficient nucleic acid detection of M. bovis. A pair of specific primers with 5' ends labeled with biotin and digoxigenin probes was designed for PCR experiments. Colloidal gold particles-labeled anti-digoxigenin IgG coated gold-labeled test strip was prepared, streptavidin was used as the detection probe, and nitrocellulose membrane coated goat anti-mouse IgG was used as the control line. Our results showed that the detection limit of the PCR-LFD was 89 fg/µL for the M. bovis DNA. The results from the test strip were highly consistent with those from real-time qPCR. This assay were highly specific for M. bovis, as there were no cross-reactions with other microorganisms tested and the detection sensitivity of the test was also relatively high (97.67%). The novel strips present a promising tool for the cost-effective and sensitive diagnosis of M. bovis.
Assuntos
Doenças dos Bovinos , Infecções por Mycoplasma , Mycoplasma bovis , Reação em Cadeia da Polimerase , Sensibilidade e Especificidade , Animais , Mycoplasma bovis/isolamento & purificação , Mycoplasma bovis/genética , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/microbiologia , Infecções por Mycoplasma/veterinária , Infecções por Mycoplasma/diagnóstico , Infecções por Mycoplasma/microbiologia , Reação em Cadeia da Polimerase/veterinária , Reação em Cadeia da Polimerase/métodos , DNA Bacteriano/análise , Coloide de Ouro/químicaRESUMO
Neosporosis is a proven disease of farm animals and dogs caused by Neospora caninum. This cross-sectional study investigates N. caninum prevalence and seroprevalence among 268 dogs. Nc5 gene PCR was carried out on dog faeces and confirmed by sequencing. Seroprevalence was detected using an indirect fluorescent antibody test (IFAT). Three age groups, gender, locality (Amman, Irbid, and Zarqa Governorates), dog type (stray, pet, and breeding), place of living (indoor/outdoor), food type (raw/cooked), having diarrhoea, having abortion in the area, and having animals nearby were tested as independent variables for associations with positivity to N. caninum using univariate and multivariable logistic regression analyses. The true prevalence of N. caninum was 34.3% (95% CI 28.4, 40.5) using the Nc5-PCR test. The true seroprevalence rate of N. caninum among dogs in Jordan was 47.9% (95% CI 41.4, 54.5) using IFAT. The sequenced isolates of Nc5-PCR products (n = 85) matched three N. caninum strains, namely, NcHareGre (n = 70, 82.4%, 95% CI 72.6-89), NC MS2 (n = 14, 16.5%, 95% CI 9.3-26.1), and L218 (n = 1, 1.2%, 95% CI 0.03-6.4). The three strains were isolated previously from three different countries and continents. N. caninum shedding is associated with abortion among dogs and animals in the area (odds ratio = 3.6). In Amman and Zarqa, living indoors reduced seroprevalence at 0.45, 0.24, and 0.02 odds ratios, respectively. Jordan shares three molecular N. caninum strains with three different countries and continents.