Detailed information on specific species of non-aureus staphylococci (NAS) has become a necessity for effective udder health control programs in South Africa. The main objective of this preliminary study was to identify the different NAS species and strains present in dairy herds in South Africa using a cost-effective method. A further objective was to investigate the effects of cow risk factors and farming systems on the NAS isolates identified. A total of 214 NAS, isolated from milk collected from 17 South African dairy herds, were identified using three diagnostic tests (API Staph test, MALDI-TOF and 16s rRNA). There was a good observed agreement between the MALDI-TOF and 16S rRNA sequencing (92.2%) and a poor observed agreement between the MALDI-TOF and API Staph (25.7%). The genetic relatedness within species was investigated in 128 of these isolates using random polymorphic amplified deoxyribonucleic acid (DNA) (RAPD), verified by multilocus sequence typing (MLST), and phylogenetic analysis and cow risk factors were investigated on species level. The main NAS species isolated were Staphylococcus chromogenes (75.2%), Staphylococcus epidermidis (9.4%) and Staphylococcus haemolyticus (8.9%). The RAPD test identified 34 Staphylococcus chromogenes, 13 Staphylococcus epidermidis and nine Staphylococcus haemolyticus strains, indicating genetic diversity amongst strains and herds. The presence of NAS intramammary infections was found to be significantly related to the farming systems, composite cow milk somatic cell count (SCC), parity and days in milk (DIM). Significantly more NAS were isolated from primiparous and from older cows. This knowledge could assist with the management of NAS on dairy farms.
{"title":"Species identification and cow risks of non-aureus staphylococci from South African dairy herds.","authors":"Inge-Marie Petzer, Christiaan Labuschagne, Lufuno Phophi, Joanne Karzis","doi":"10.4102/ojvr.v89i1.2021","DOIUrl":"https://doi.org/10.4102/ojvr.v89i1.2021","url":null,"abstract":"<p><p>Detailed information on specific species of non-aureus staphylococci (NAS) has become a necessity for effective udder health control programs in South Africa. The main objective of this preliminary study was to identify the different NAS species and strains present in dairy herds in South Africa using a cost-effective method. A further objective was to investigate the effects of cow risk factors and farming systems on the NAS isolates identified. A total of 214 NAS, isolated from milk collected from 17 South African dairy herds, were identified using three diagnostic tests (API Staph test, MALDI-TOF and 16s rRNA). There was a good observed agreement between the MALDI-TOF and 16S rRNA sequencing (92.2%) and a poor observed agreement between the MALDI-TOF and API Staph (25.7%). The genetic relatedness within species was investigated in 128 of these isolates using random polymorphic amplified deoxyribonucleic acid (DNA) (RAPD), verified by multilocus sequence typing (MLST), and phylogenetic analysis and cow risk factors were investigated on species level. The main NAS species isolated were Staphylococcus chromogenes (75.2%), Staphylococcus epidermidis (9.4%) and Staphylococcus haemolyticus (8.9%). The RAPD test identified 34 Staphylococcus chromogenes, 13 Staphylococcus epidermidis and nine Staphylococcus haemolyticus strains, indicating genetic diversity amongst strains and herds. The presence of NAS intramammary infections was found to be significantly related to the farming systems, composite cow milk somatic cell count (SCC), parity and days in milk (DIM). Significantly more NAS were isolated from primiparous and from older cows. This knowledge could assist with the management of NAS on dairy farms.</p>","PeriodicalId":54685,"journal":{"name":"Onderstepoort Journal of Veterinary Research","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2022-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9350540/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40580616","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sara Locke, Vinny Naidoo, Ibrahim Hassan, Neil Duncan
Diclofenac was responsible for the decimation of Gyps vulture species on the Indian subcontinent during the 1980s and 1990s. Gyps vultures are extremely sensitive (the lethal dose 50 [LD50] ~ 0.1 mg/kg - 0.2 mg/kg), with toxicity appearing to be linked to metabolic deficiency, demonstrated by the long T1/2 (~12 h - 17 h). This is in striking comparison to the domestic chicken (Gallus gallus domesticus), in which the LD50 is ~10 mg/kg and the T1/2 is ~1 h. The phase 1 cytochrome P450 (CYP) 2C subfamily has been cited as a possible reason for metabolic deficiency. The aim of this study was to determine if CYP2C9 homolog pharmacogenomic differences amongst avian species is driving diclofenac toxicity in Gyps vultures. We exposed each of 10 CYP-inhibited test group chickens to a unique dose of diclofenac (as per the Organisation for Economic Co-operation and Development [OECD] toxicity testing guidelines) and compared the toxicity and pharmacokinetic results to control group birds that received no CYP inhibitor. Although no differences were noted in the LD50 values for each group (11.92 mg/kg in the CYP-inhibited test group and 11.58 mg/kg in the control group), the pharmacokinetic profile of the test group was suggestive of partial inhibition of CYP metabolism. Evaluation of the metabolite peaks produced also suggested partial metabolic inhibition in test group birds, as they produced lower amounts of metabolites for one of the three peaks demonstrated and had higher diclofenac exposure. This pilot study supports the hypothesis that CYP metabolism is varied amongst bird species and may explain the higher resilience to diclofenac in the chicken versus vultures.
{"title":"Effect of cytochrome P450 inhibition on toxicity of diclofenac in chickens: Unravelling toxicity in Gyps vultures.","authors":"Sara Locke, Vinny Naidoo, Ibrahim Hassan, Neil Duncan","doi":"10.4102/ojvr.v89i1.1978","DOIUrl":"https://doi.org/10.4102/ojvr.v89i1.1978","url":null,"abstract":"<p><p>Diclofenac was responsible for the decimation of Gyps vulture species on the Indian subcontinent during the 1980s and 1990s. Gyps vultures are extremely sensitive (the lethal dose 50 [LD50] ~ 0.1 mg/kg - 0.2 mg/kg), with toxicity appearing to be linked to metabolic deficiency, demonstrated by the long T1/2 (~12 h - 17 h). This is in striking comparison to the domestic chicken (Gallus gallus domesticus), in which the LD50 is ~10 mg/kg and the T1/2 is ~1 h. The phase 1 cytochrome P450 (CYP) 2C subfamily has been cited as a possible reason for metabolic deficiency. The aim of this study was to determine if CYP2C9 homolog pharmacogenomic differences amongst avian species is driving diclofenac toxicity in Gyps vultures. We exposed each of 10 CYP-inhibited test group chickens to a unique dose of diclofenac (as per the Organisation for Economic Co-operation and Development [OECD] toxicity testing guidelines) and compared the toxicity and pharmacokinetic results to control group birds that received no CYP inhibitor. Although no differences were noted in the LD50 values for each group (11.92 mg/kg in the CYP-inhibited test group and 11.58 mg/kg in the control group), the pharmacokinetic profile of the test group was suggestive of partial inhibition of CYP metabolism. Evaluation of the metabolite peaks produced also suggested partial metabolic inhibition in test group birds, as they produced lower amounts of metabolites for one of the three peaks demonstrated and had higher diclofenac exposure. This pilot study supports the hypothesis that CYP metabolism is varied amongst bird species and may explain the higher resilience to diclofenac in the chicken versus vultures.</p>","PeriodicalId":54685,"journal":{"name":"Onderstepoort Journal of Veterinary Research","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2022-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9257893/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40574799","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-03-01DOI: 10.2478/jvetres-2022-0007
Huanan Gao, Jinping Wang, Z. Yang, Jiarui Xie, Yuwen He, Q. Hong, A. Xin
Abstract Introduction Akabane virus (AKAV) has been detected in a variety of host species in China, but there are only limited records of its occurrence in goats. However, more attention needs to be paid to understanding the diversity of viruses in this species. The aim of the study was to explore the genotype characteristics and variation trend of AKAV and their relationship with virulence in Yunnan, China. Material and Methods Blood samples were collected from goats during routine surveillance of goat diseases in Yunnan province in 2019. The AKAV CX-01 strain was isolated using BHK-21 cells. To understand pathogenicity, the virus was intraperitoneally (IP) and intracerebrally (IC) inoculated into suckling mice and tissue samples were subsequently analysed histopathologically and immunohistochemically. Results Akabane virus CX-01 strain induced encephalitis and impairment of the central nervous system with fatal consequences. Phylogenetic analysis based on the ORF sequences of the small segments indicated that the AKAV isolate used was most closely related to the GD18134/2018 Chinese midge and bovine NM BS/1strains, while phylogenetic analysis based on the medium segments showed a close relationship between CX-01 and the Chinese GLXCH01 strain. Conclusion The CX-01 isolate was related to AKAV genogroup Ia and probably originated from a recombination of different strains.
{"title":"Genetic and Pathogenic Characterisation of a Virulent Akabane Virus Isolated from Goats in Yunnan, China","authors":"Huanan Gao, Jinping Wang, Z. Yang, Jiarui Xie, Yuwen He, Q. Hong, A. Xin","doi":"10.2478/jvetres-2022-0007","DOIUrl":"https://doi.org/10.2478/jvetres-2022-0007","url":null,"abstract":"Abstract Introduction Akabane virus (AKAV) has been detected in a variety of host species in China, but there are only limited records of its occurrence in goats. However, more attention needs to be paid to understanding the diversity of viruses in this species. The aim of the study was to explore the genotype characteristics and variation trend of AKAV and their relationship with virulence in Yunnan, China. Material and Methods Blood samples were collected from goats during routine surveillance of goat diseases in Yunnan province in 2019. The AKAV CX-01 strain was isolated using BHK-21 cells. To understand pathogenicity, the virus was intraperitoneally (IP) and intracerebrally (IC) inoculated into suckling mice and tissue samples were subsequently analysed histopathologically and immunohistochemically. Results Akabane virus CX-01 strain induced encephalitis and impairment of the central nervous system with fatal consequences. Phylogenetic analysis based on the ORF sequences of the small segments indicated that the AKAV isolate used was most closely related to the GD18134/2018 Chinese midge and bovine NM BS/1strains, while phylogenetic analysis based on the medium segments showed a close relationship between CX-01 and the Chinese GLXCH01 strain. Conclusion The CX-01 isolate was related to AKAV genogroup Ia and probably originated from a recombination of different strains.","PeriodicalId":54685,"journal":{"name":"Onderstepoort Journal of Veterinary Research","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2022-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73174461","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-03-01DOI: 10.2478/jvetres-2022-0013
K. Urbaniak, A. Kowalczyk, M. Pomorska-Mól, K. Kwit, I. Markowska-Daniel
Abstract Introduction The lack of proofreading activity of the viral polymerase and the segmented nature of the influenza A virus (IAV) genome are responsible for the genetic diversity of IAVs and for their ability to adapt to a new host. We tried to adapt avian IAV (avIAV) to the pig by serial passages in vivo and assessed the occurrence of point mutations and their influence on viral fitness in the pig’s body. Material and Methods A total of 25 in vivo avIAV passages of the A/duck/Bavaria/77 strain were performed by inoculation of 50 piglets, and after predetermined numbers of passages 20 uninoculated piglets were exposed to the virus through contact with inoculated animals. Clinical signs of swine influenza were assessed daily. Nasal swabs and lung tissue were used to detect IAV RNA by real-time RT-PCR and isolates from selected passages were sequenced. Results Apart from a rise in rectal temperature and a sporadic cough, no typical clinical signs were observed in infected pigs. The original strain required 20 passages to improve its replication ability noticeably. A total of 29 amino-acid substitutions were identified. Eighteen of them were detected in the first sequenced isolate, of which 16 were also in all other analysed strains. Additional mutations were detected with more passages. One substitution, threonine (T) 135 to serine (S) in neuraminidase (NA), was only detected in an IAV isolate from a contact-exposed piglet. Conclusion Passaging 25 times allowed us to obtain a partially swine-adapted IAV. The improvement in isolate replication ability was most likely related to S654 to glycine (G) substitution in the basic protein (PB) 1 as well as to aspartic acid (D) 701 to asparagine (N) and arginine (R) 477 to G in PB2, glutamic acid (E) 204 to D and G239E in haemagglutinin and T135S in NA.
{"title":"Effect of Serial in Vivo Passages on The Adaptation of H1N1 Avian Influenza Virus To Pigs","authors":"K. Urbaniak, A. Kowalczyk, M. Pomorska-Mól, K. Kwit, I. Markowska-Daniel","doi":"10.2478/jvetres-2022-0013","DOIUrl":"https://doi.org/10.2478/jvetres-2022-0013","url":null,"abstract":"Abstract Introduction The lack of proofreading activity of the viral polymerase and the segmented nature of the influenza A virus (IAV) genome are responsible for the genetic diversity of IAVs and for their ability to adapt to a new host. We tried to adapt avian IAV (avIAV) to the pig by serial passages in vivo and assessed the occurrence of point mutations and their influence on viral fitness in the pig’s body. Material and Methods A total of 25 in vivo avIAV passages of the A/duck/Bavaria/77 strain were performed by inoculation of 50 piglets, and after predetermined numbers of passages 20 uninoculated piglets were exposed to the virus through contact with inoculated animals. Clinical signs of swine influenza were assessed daily. Nasal swabs and lung tissue were used to detect IAV RNA by real-time RT-PCR and isolates from selected passages were sequenced. Results Apart from a rise in rectal temperature and a sporadic cough, no typical clinical signs were observed in infected pigs. The original strain required 20 passages to improve its replication ability noticeably. A total of 29 amino-acid substitutions were identified. Eighteen of them were detected in the first sequenced isolate, of which 16 were also in all other analysed strains. Additional mutations were detected with more passages. One substitution, threonine (T) 135 to serine (S) in neuraminidase (NA), was only detected in an IAV isolate from a contact-exposed piglet. Conclusion Passaging 25 times allowed us to obtain a partially swine-adapted IAV. The improvement in isolate replication ability was most likely related to S654 to glycine (G) substitution in the basic protein (PB) 1 as well as to aspartic acid (D) 701 to asparagine (N) and arginine (R) 477 to G in PB2, glutamic acid (E) 204 to D and G239E in haemagglutinin and T135S in NA.","PeriodicalId":54685,"journal":{"name":"Onderstepoort Journal of Veterinary Research","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2022-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81556388","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-03-01DOI: 10.2478/jvetres-2022-0017
K. Śmietanka, Edyta Świętoń, K. Wyrostek, E. Kozak, K. Tarasiuk, Natalia Styś-Fijoł, Kamila Dziadek, K. Niemczuk
Abstract Introduction Highly pathogenic avian influenza (HPAI) outbreaks caused by the Gs/Gd lineage of H5Nx viruses occur in Poland with increased frequency. The article provides an update on the HPAI situation in the 2020/2021 season and studies the possible factors that caused the exceptionally fast spread of the virus. Material and Methods Samples from poultry and wild birds delivered for HPAI diagnosis were tested by real-time RT-PCR and a representative number of detected viruses were submitted for partial or full-genome characterisation. Information yielded by veterinary inspection was used for descriptive analysis of the epidemiological situation. Results The scale of the epidemic in the 2020/2021 season was unprecedented in terms of duration (November 2020–August 2021), number of outbreaks in poultry (n = 357), wild bird events (n = 92) and total number of affected domestic birds (approximately ~14 million). The major drivers of the virus spread were the harsh winter conditions in February 2020 followed by the introduction of the virus to high-density poultry areas in March 2021. All tested viruses belonged to H5 clade 2.3.4.4b with significant intra-clade diversity and in some cases clearly distinguished clusters. Conclusion The HPAI epidemic in 2020/2021 in Poland struck with unprecedented force. The conventional control measures may have limited effectiveness to break the transmission chain in areas with high concentrations of poultry.
{"title":"Highly Pathogenic Avian Influenza H5Nx in Poland in 2020/2021: a Descriptive Epidemiological Study of a Large-scale Epidemic","authors":"K. Śmietanka, Edyta Świętoń, K. Wyrostek, E. Kozak, K. Tarasiuk, Natalia Styś-Fijoł, Kamila Dziadek, K. Niemczuk","doi":"10.2478/jvetres-2022-0017","DOIUrl":"https://doi.org/10.2478/jvetres-2022-0017","url":null,"abstract":"Abstract Introduction Highly pathogenic avian influenza (HPAI) outbreaks caused by the Gs/Gd lineage of H5Nx viruses occur in Poland with increased frequency. The article provides an update on the HPAI situation in the 2020/2021 season and studies the possible factors that caused the exceptionally fast spread of the virus. Material and Methods Samples from poultry and wild birds delivered for HPAI diagnosis were tested by real-time RT-PCR and a representative number of detected viruses were submitted for partial or full-genome characterisation. Information yielded by veterinary inspection was used for descriptive analysis of the epidemiological situation. Results The scale of the epidemic in the 2020/2021 season was unprecedented in terms of duration (November 2020–August 2021), number of outbreaks in poultry (n = 357), wild bird events (n = 92) and total number of affected domestic birds (approximately ~14 million). The major drivers of the virus spread were the harsh winter conditions in February 2020 followed by the introduction of the virus to high-density poultry areas in March 2021. All tested viruses belonged to H5 clade 2.3.4.4b with significant intra-clade diversity and in some cases clearly distinguished clusters. Conclusion The HPAI epidemic in 2020/2021 in Poland struck with unprecedented force. The conventional control measures may have limited effectiveness to break the transmission chain in areas with high concentrations of poultry.","PeriodicalId":54685,"journal":{"name":"Onderstepoort Journal of Veterinary Research","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2022-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89182940","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-03-01DOI: 10.2478/jvetres-2022-0016
M. Walczak, M. Juszkiewicz, K. Szymankiewicz, A. Szczotka-Bochniarz, G. Woźniakowski
Abstract Introduction African swine fever virus (ASFV) causes one of the most dangerous diseases of pigs and wild boar – African swine fever (ASF). Since its second introduction into Europe (in 2007), the disease has been spreading consistently, and now ASF-free European countries are at risk. Complex interactions between the host’s immune system and the virus have long prevented the development of a safe vaccine against ASF. This study analysed the possibility of neutralisation of the ASFV in vitro by sera collected from ASF-survivor animals. Material and Methods Two pig and three wild boar serum samples were collected from previously selected potential ASF survivors. All sera presented high antibody titres (>5 log10/mL). Primary alveolar macrophages were cultured in growth medium containing 10% and 20% concentrations of selected sera and infected with a haemadsorbing ASFV strain (Pol18_28298_O111, genotype II). The progress of infection was investigated under a light microscope by observing the cytopathic effect (CPE) and the haemadsorption phenomenon. Growth kinetics were investigated using a real-time PCR assay. Results Haemadsorption inhibition was detected in the presence of almost all selected sera; however, the inhibition of virus replication in vitro was excluded. In all samples, a CPE and decreasing quantification cycle values of the viral DNA were found. Conclusion Anti-ASFV antibodies alone are not able to inhibit virus replication. Interactions between the humoral and cellular immune response which effectively combat the disease are implicated in an ASF-survivor’s organism.
{"title":"ASF -survivors’ Sera Do Not Inhibit African Swine Fever Virus Replication in Vitro","authors":"M. Walczak, M. Juszkiewicz, K. Szymankiewicz, A. Szczotka-Bochniarz, G. Woźniakowski","doi":"10.2478/jvetres-2022-0016","DOIUrl":"https://doi.org/10.2478/jvetres-2022-0016","url":null,"abstract":"Abstract Introduction African swine fever virus (ASFV) causes one of the most dangerous diseases of pigs and wild boar – African swine fever (ASF). Since its second introduction into Europe (in 2007), the disease has been spreading consistently, and now ASF-free European countries are at risk. Complex interactions between the host’s immune system and the virus have long prevented the development of a safe vaccine against ASF. This study analysed the possibility of neutralisation of the ASFV in vitro by sera collected from ASF-survivor animals. Material and Methods Two pig and three wild boar serum samples were collected from previously selected potential ASF survivors. All sera presented high antibody titres (>5 log10/mL). Primary alveolar macrophages were cultured in growth medium containing 10% and 20% concentrations of selected sera and infected with a haemadsorbing ASFV strain (Pol18_28298_O111, genotype II). The progress of infection was investigated under a light microscope by observing the cytopathic effect (CPE) and the haemadsorption phenomenon. Growth kinetics were investigated using a real-time PCR assay. Results Haemadsorption inhibition was detected in the presence of almost all selected sera; however, the inhibition of virus replication in vitro was excluded. In all samples, a CPE and decreasing quantification cycle values of the viral DNA were found. Conclusion Anti-ASFV antibodies alone are not able to inhibit virus replication. Interactions between the humoral and cellular immune response which effectively combat the disease are implicated in an ASF-survivor’s organism.","PeriodicalId":54685,"journal":{"name":"Onderstepoort Journal of Veterinary Research","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2022-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86962450","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-03-01DOI: 10.2478/jvetres-2022-0005
Y. Badr, M. M. Rahman, Y. Ohno, Keita Ishijima, Ken Maeda, K. Kohyama, Y. Kamatari, K. Shimizu, Ayaka Okada, Y. Inoshima
Abstract Introduction Seal parapoxvirus (SPPV) infection has been reported among pinnipeds in aquaria in Japan; however, its seroprevalence is unknown. Therefore, an enzyme-linked immunosorbent assay (ELISA) was developed for serological diagnosis of SPPV infection. Material and Methods The gene encoding the major envelope protein of SPPV was cloned into the eukaryotic expression vector pAcGFP1-N1, which encodes the green fluorescence protein (GFP), thereby producing a fusion protein (Env-GFP). Parental and cloned vector DNA was independently transfected into cultured seal cells for the expression of GFP and Env-GFP. The wells of an ELISA plate were coated with either GFP- or Env-GFP-transfected cell lysates. The light absorbance of each serum sample was adjusted by subtracting the absorbance of GFP-coated wells from that of Env-GFP-coated wells. Sera from two spotted seals (Phoca largha), six beluga whales (Delphinapterus leucas), three Pacific white-sided dolphins (Lagenorhynchus obliquidens), and ten bottlenose dolphins (Tursiops truncatus) from an aquarium in Japan were examined using the ELISA. Results Positive reactions were not observed, except in one preserved sample collected ten years ago from a naturally SPPV-infected spotted seal. Conclusion The established ELISA could be useful in screening marine mammal sera for anti-SPPV antibodies.
{"title":"A New Enzyme-linked Immunosorbent Assay for Serological Diagnosis of Seal Parapoxvirus Infection in Marine Mammals","authors":"Y. Badr, M. M. Rahman, Y. Ohno, Keita Ishijima, Ken Maeda, K. Kohyama, Y. Kamatari, K. Shimizu, Ayaka Okada, Y. Inoshima","doi":"10.2478/jvetres-2022-0005","DOIUrl":"https://doi.org/10.2478/jvetres-2022-0005","url":null,"abstract":"Abstract Introduction Seal parapoxvirus (SPPV) infection has been reported among pinnipeds in aquaria in Japan; however, its seroprevalence is unknown. Therefore, an enzyme-linked immunosorbent assay (ELISA) was developed for serological diagnosis of SPPV infection. Material and Methods The gene encoding the major envelope protein of SPPV was cloned into the eukaryotic expression vector pAcGFP1-N1, which encodes the green fluorescence protein (GFP), thereby producing a fusion protein (Env-GFP). Parental and cloned vector DNA was independently transfected into cultured seal cells for the expression of GFP and Env-GFP. The wells of an ELISA plate were coated with either GFP- or Env-GFP-transfected cell lysates. The light absorbance of each serum sample was adjusted by subtracting the absorbance of GFP-coated wells from that of Env-GFP-coated wells. Sera from two spotted seals (Phoca largha), six beluga whales (Delphinapterus leucas), three Pacific white-sided dolphins (Lagenorhynchus obliquidens), and ten bottlenose dolphins (Tursiops truncatus) from an aquarium in Japan were examined using the ELISA. Results Positive reactions were not observed, except in one preserved sample collected ten years ago from a naturally SPPV-infected spotted seal. Conclusion The established ELISA could be useful in screening marine mammal sera for anti-SPPV antibodies.","PeriodicalId":54685,"journal":{"name":"Onderstepoort Journal of Veterinary Research","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2022-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90304308","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract Introduction Hypoxia is a common pathological condition after spinal cord injury. Oestrogen-related receptor alpha (ERRα), as a key regulator of energy metabolism and mitochondrial functions, plays an important role in maintaining cell homeostasis. However, its role in hypoxic spinal microglia has not been fully elaborated. This study investigated the receptor’s activity when these cells are hypoxic and used as an in vitro model. Material and Methods In this study, microglia (BV2) were exposed to cobalt chloride as a hypoxic model, and the inverse agonist of ERRα, XCT790, and pyrido[1,2-α]-pyrimidin-4-one were used to regulate the expression of the receptor to explore the ERRα-related mechanisms involved in hypoxic spinal cord injury (SCI). Results ERRα promoted autophagy in BV2 cells and inhibited the activation of the p38 mitogen-activated protein kinase (MAPK) pathway and the expression of anti-inflammatory factors under hypoxic conditions. It also promoted the expression of fibronectin type III domain containing protein 5 (FNDC5). Conclusion When a hypoxic SCI occurs, ERRα may maintain the homeostasis of spinal cord nerve cells by regulating autophagy and the p38MAPK/nuclear factor-kappa B cell and FNDC5/brain-derived neurotrophic factor signalling pathways, which are beneficial to the recovery of these cells.
{"title":"Estrogen-related Receptor α (ERRα) Functions in The Hypoxic Injury of Microglial Cells","authors":"Chaowei Deng, Tingting Zhu, S. Lian, Jianfa Wang, Rui Wu, Jia-san Zheng","doi":"10.2478/jvetres-2022-0009","DOIUrl":"https://doi.org/10.2478/jvetres-2022-0009","url":null,"abstract":"Abstract Introduction Hypoxia is a common pathological condition after spinal cord injury. Oestrogen-related receptor alpha (ERRα), as a key regulator of energy metabolism and mitochondrial functions, plays an important role in maintaining cell homeostasis. However, its role in hypoxic spinal microglia has not been fully elaborated. This study investigated the receptor’s activity when these cells are hypoxic and used as an in vitro model. Material and Methods In this study, microglia (BV2) were exposed to cobalt chloride as a hypoxic model, and the inverse agonist of ERRα, XCT790, and pyrido[1,2-α]-pyrimidin-4-one were used to regulate the expression of the receptor to explore the ERRα-related mechanisms involved in hypoxic spinal cord injury (SCI). Results ERRα promoted autophagy in BV2 cells and inhibited the activation of the p38 mitogen-activated protein kinase (MAPK) pathway and the expression of anti-inflammatory factors under hypoxic conditions. It also promoted the expression of fibronectin type III domain containing protein 5 (FNDC5). Conclusion When a hypoxic SCI occurs, ERRα may maintain the homeostasis of spinal cord nerve cells by regulating autophagy and the p38MAPK/nuclear factor-kappa B cell and FNDC5/brain-derived neurotrophic factor signalling pathways, which are beneficial to the recovery of these cells.","PeriodicalId":54685,"journal":{"name":"Onderstepoort Journal of Veterinary Research","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2022-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91003076","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-03-01DOI: 10.2478/jvetres-2022-0008
T. Kotnik, A. V. Rataj, B. Šoba
Abstract Introduction The prevalence of Dirofilaria repens in dogs in countries bordering Slovenia ranges from 1.5% to 47.3%. The aim of this study was to estimate its prevalence in Slovenian dogs and to present the cases of dirofilariasis diagnosed in humans from 2010 to 2020. Material and Methods Epidemiological data were collected and blood samples were taken from 465 dogs older than one year and born in Slovenia. A real-time PCR was performed on all samples to detect filarioid DNA, and a D. repens-and D. immitis-specific real-time PCR was performed on positive samples. Blood samples from 446 dogs were tested for Dirofilaria spp. using a modified Knott’s test. Human cases were diagnosed from histological sections of excised subcutaneous nodules. Descriptive statistics were used to characterise the samples. The one-sample nonparametric chi-squared test was used to assess whether categories of a variable were equally distributed. Results Three dogs’ samples tested positive for D. repens using the species-specific real-time PCR, while D. immitis DNA was not detected. The modified Knott’s test was positive in two of the three PCR-positive dogs, two of which had never travelled outside Slovenia’s borders. Four human patients with D. repens dirofilariasis were diagnosed. Since their travel history was unknown, autochthonous transmission could not be confirmed. Conclusion Our study demonstrated a 0.64% prevalence of D. repens infection in dogs in Slovenia. Two cases could be autochthonous.
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Pub Date : 2022-03-01DOI: 10.2478/jvetres-2022-0006
L. Perillo, G. Cascone, F. Antoci, G. Piccione, C. Giannetto, Rosario Salonia, F. Salina, E. Giudice, V. Monteverde, F. Licitra
Abstract Introduction The aim of the study was to investigate the relationship between biosecurity as scored on the Italian National Animal Welfare Reference Centre (Centro di Referenza Nazionale per il Benessere Animale – CReNBA) checklist and the prevalence of Mycobacterium avium subsp. paratuberculosis, Chlamydophila abortus and Neospora caninum on dairy farms located in Ragusa, Italy. Material and Methods The checklist was used to assign an animal welfare score to 31 dairy farms. Twenty-one farms with a moderate score (>33%, <66%) formed group 1, and 10 farms with a high score (>66%) were group 2. Blood samples were collected from all cows on each farm to investigate the titres of antibodies against the relevant pathogens. Two-way analysis of variance was applied to assess differences between the two experimental groups and the Mann–Whitney test was applied to evaluate prevalence differences in the tested parasites between the groups. Results All tested farms had a score that classified them as either good or excellent. A higher incidence of Neospora caninum was observed in group 1. The incidences of the other two parasites were no different between the two groups. Conclusion The CReNBA checklist represents an impartial, reproducible, functional and smart instrument based on risk analysis and assigns a farm a mathematical animal welfare score. Among the parasites tested for, only Neospora caninum had prevalence influenced by biosecurity. Our preliminary results highlighted the positive associations between good animal welfare, high levels of biosecurity, and the prevention of the infectious diseases caused by the parasites in our focus, which are common on dairy farms.
摘要简介本研究的目的是调查意大利国家动物福利参考中心(Centro di Referenza Nazionale per il Benessere Animale - CReNBA)检查表中生物安全性得分与鸟分枝杆菌亚种流行率之间的关系。在意大利拉古萨的奶牛场发现了副结核、流产衣原体和犬新孢子虫。材料与方法采用检查表对31家奶牛场进行动物福利评分。21个中等得分的农场(>33%，66%)为第2组。从每个农场的所有奶牛中采集血样，以调查针对相关病原体的抗体滴度。采用双向方差分析评价两实验组间的差异，采用Mann-Whitney检验评价两组间被测寄生虫患病率差异。结果所有受测农场的得分均为良好或优秀。第1组犬新孢子虫发病率较高。另外两种寄生虫的发病率在两组之间没有差异。结论CReNBA清单代表了一种基于风险分析的公正、可复制、功能和智能的工具，并为农场分配了一个数学动物福利评分。在检测的寄生虫中，只有犬新孢子虫受生物安全因素的影响。我们的初步结果强调了良好的动物福利、高水平的生物安全与预防由我们关注的寄生虫引起的传染病之间的正相关关系，这些疾病在奶牛场很常见。
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