Pub Date : 2025-02-01DOI: 10.1016/j.vetmic.2024.110343
Caiying Wang , Yue Zhang , Shanshan Yang , Huub F.J. Savelkoul , Christine A. Jansen , Guangliang Liu
Porcine epidemic diarrhea virus (PEDV) is a coronavirus that induces diarrhea in pigs, leading to severe economic losses in the global pig industry. Currently, effective antiviral treatments for porcine epidemic diarrhea (PED) are rarely available for clinical use. Zinc (Zn2+), an essential mineral, is known to reduce diarrhea in piglets transitioning from milk to solid feed by modulating immune system activity. In this study, the role of Zn2+ in regulating PEDV infection was investigated to explore its potential for reducing diarrhea. Our findings show that Zn2+ inhibits PEDV replication in Vero-E6 cells by inducing autophagy. Notably, we demonstrated that autophagy negatively regulates PEDV infection, as confirmed by the use of autophagy inhibitor (3-MA) and activator (RAPA). Further analysis revealed that PEDV infection activates the Akt-mTOR signaling pathway, while Zn2+ inhibits this pathway in Vero-E6 cells. Additionally, overexpression of Akt and AktSer473 plasmids in Vero-E6 cells highlights the role of Akt phosphorylation in the Zn2+ induced autophagy that inhibits PEDV replication. In summary, this study identifies a mechanism by which Zn2+ suppresses PEDV infection through the Akt-mTOR pathway by mediating autophagy. These findings provide valuable insights into the potential use of Zn2+ as an effective antiviral agent in vivo.
{"title":"Zn2+ inhibits PEDV replication by inducing autophagy through the Akt-mTOR pathway","authors":"Caiying Wang , Yue Zhang , Shanshan Yang , Huub F.J. Savelkoul , Christine A. Jansen , Guangliang Liu","doi":"10.1016/j.vetmic.2024.110343","DOIUrl":"10.1016/j.vetmic.2024.110343","url":null,"abstract":"<div><div>Porcine epidemic diarrhea virus (PEDV) is a coronavirus that induces diarrhea in pigs, leading to severe economic losses in the global pig industry. Currently, effective antiviral treatments for porcine epidemic diarrhea (PED) are rarely available for clinical use. Zinc (Zn<sup>2+</sup>), an essential mineral, is known to reduce diarrhea in piglets transitioning from milk to solid feed by modulating immune system activity. In this study, the role of Zn<sup>2+</sup> in regulating PEDV infection was investigated to explore its potential for reducing diarrhea. Our findings show that Zn<sup>2+</sup> inhibits PEDV replication in Vero-E6 cells by inducing autophagy. Notably, we demonstrated that autophagy negatively regulates PEDV infection, as confirmed by the use of autophagy inhibitor (3-MA) and activator (RAPA). Further analysis revealed that PEDV infection activates the Akt-mTOR signaling pathway, while Zn<sup>2+</sup> inhibits this pathway in Vero-E6 cells. Additionally, overexpression of Akt and Akt<sub>Ser473</sub> plasmids in Vero-E6 cells highlights the role of Akt phosphorylation in the Zn<sup>2+</sup> induced autophagy that inhibits PEDV replication. In summary, this study identifies a mechanism by which Zn<sup>2+</sup> suppresses PEDV infection through the Akt-mTOR pathway by mediating autophagy. These findings provide valuable insights into the potential use of Zn<sup>2+</sup> as an effective antiviral agent <em>in vivo</em>.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"301 ","pages":"Article 110343"},"PeriodicalIF":2.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142872177","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 : 2025-02-01DOI: 10.1016/j.vetmic.2024.110345
Qingqing Gao, Qianlong Xing, Yunyan Sun, Zhengliang Li, Song Gao
Avian pathogenic Escherichia coli (APEC) constitutes a significant threat to poultry health worldwide, causing colibacillosis and inflicting substantial economic losses. The ability to resist serum-mediated killing is a key virulence factor enabling APEC to circumvent the host immune system and establish systemic infection. In this study, we employed mariner-based transposon mutagenesis to generate a mutant library of APEC strain E058 and screened for mutants with reduced serum resistance. We identified a transposon insertion within the sspB gene of the sspA-sspB operon that conferred significantly reduced serum resistance. Targeted gene knockout experiments confirmed that both sspA and sspB contribute to serum resistance, with the double mutant (ΔsspAΔsspB) displaying a more pronounced susceptibility to serum compared to the single gene knockouts (ΔsspA and ΔsspB). Furthermore, in vivo challenge experiments in chickens demonstrated that disruption of the sspA-sspB operon significantly attenuated APEC virulence. Our study also reveals that the sspA-sspB operon plays a role in biofilm formation and promotes intracellular survival within macrophages, suggesting a multifaceted contribution to APEC pathogenesis. These findings highlight the sspA-sspB operon as a promising target for the development of novel therapeutics against APEC infections in poultry.
{"title":"Transposon mutagenesis identifies the sspA-sspB operon as essential for serum resistance and virulence in avian pathogenic Escherichia coli","authors":"Qingqing Gao, Qianlong Xing, Yunyan Sun, Zhengliang Li, Song Gao","doi":"10.1016/j.vetmic.2024.110345","DOIUrl":"10.1016/j.vetmic.2024.110345","url":null,"abstract":"<div><div>Avian pathogenic <em>Escherichia coli</em> (APEC) constitutes a significant threat to poultry health worldwide, causing colibacillosis and inflicting substantial economic losses. The ability to resist serum-mediated killing is a key virulence factor enabling APEC to circumvent the host immune system and establish systemic infection. In this study, we employed mariner-based transposon mutagenesis to generate a mutant library of APEC strain E058 and screened for mutants with reduced serum resistance. We identified a transposon insertion within the <em>sspB</em> gene of the <em>sspA</em>-<em>sspB</em> operon that conferred significantly reduced serum resistance. Targeted gene knockout experiments confirmed that both <em>sspA</em> and <em>sspB</em> contribute to serum resistance, with the double mutant (Δ<em>sspA</em>Δ<em>sspB</em>) displaying a more pronounced susceptibility to serum compared to the single gene knockouts (Δ<em>sspA</em> and Δ<em>sspB</em>). Furthermore, <em>in vivo</em> challenge experiments in chickens demonstrated that disruption of the <em>sspA</em>-<em>sspB</em> operon significantly attenuated APEC virulence. Our study also reveals that the <em>sspA</em>-<em>sspB</em> operon plays a role in biofilm formation and promotes intracellular survival within macrophages, suggesting a multifaceted contribution to APEC pathogenesis. These findings highlight the <em>sspA</em>-<em>sspB</em> operon as a promising target for the development of novel therapeutics against APEC infections in poultry.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"301 ","pages":"Article 110345"},"PeriodicalIF":2.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142898451","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 : 2025-02-01DOI: 10.1016/j.vetmic.2024.110359
Zixin Li , Yudi Pan , Yanxiang Zhou , Jianshuang Cui , Hailiang Ge , Wei Zhao , Li Feng , Jin Tian
Rotavirus Group A (RVA) is a primary pathogen that causes viral diarrhea in humans and animals. Porcine rotaviruses (PoRVs) are widely epidemic in pig farms in China, causing great economic losses to the swine industry. In the past 30 years, the G5 RVA had been the main epidemic genotype in pig farms worldwide. However, G9 RVA is an emerging genotype that is gradually becoming prevalent in humans and animals. To explore its potential mechanism, we isolated G9P[23] and G5P[23] rotaviruses, named 923 H and NG523 respectively, from diarrheal samples and compared the growth curves and virulence of two strains. In vitro experiments revealed that pig small intestine epithelial cells were more susceptible to 923 H strain. In vivo experiments showed that 923 H strain was more virulent than NG523 strain, causing more severe damage to piglets. The viral load of G9 infection groups in intestinal and extra-intestinal tissues was higher than that of G5 infection group. Histopathological examination showed cell degeneration, necrosis and nuclear condensation in the jejunum of G9 RVA infection group as well as more inflammatory cell infiltration and tissue destruction in the lung of G9 RVA infection group. Our results indicate that 923 H strain is more pathogenic than NG523 strain, which provides new insights into the widespread epidemic of G9 RVA in pig farms.
{"title":"Pathogenicity comparison between porcine G9P[23] and G5P[23] RVA in piglets","authors":"Zixin Li , Yudi Pan , Yanxiang Zhou , Jianshuang Cui , Hailiang Ge , Wei Zhao , Li Feng , Jin Tian","doi":"10.1016/j.vetmic.2024.110359","DOIUrl":"10.1016/j.vetmic.2024.110359","url":null,"abstract":"<div><div>Rotavirus Group A (RVA) is a primary pathogen that causes viral diarrhea in humans and animals. Porcine rotaviruses (PoRVs) are widely epidemic in pig farms in China, causing great economic losses to the swine industry. In the past 30 years, the G5 RVA had been the main epidemic genotype in pig farms worldwide. However, G9 RVA is an emerging genotype that is gradually becoming prevalent in humans and animals. To explore its potential mechanism, we isolated G9P[23] and G5P[23] rotaviruses, named 923 H and NG523 respectively, from diarrheal samples and compared the growth curves and virulence of two strains. In vitro experiments revealed that pig small intestine epithelial cells were more susceptible to 923 H strain. In vivo experiments showed that 923 H strain was more virulent than NG523 strain, causing more severe damage to piglets. The viral load of G9 infection groups in intestinal and extra-intestinal tissues was higher than that of G5 infection group. Histopathological examination showed cell degeneration, necrosis and nuclear condensation in the jejunum of G9 RVA infection group as well as more inflammatory cell infiltration and tissue destruction in the lung of G9 RVA infection group. Our results indicate that 923 H strain is more pathogenic than NG523 strain, which provides new insights into the widespread epidemic of G9 RVA in pig farms.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"301 ","pages":"Article 110359"},"PeriodicalIF":2.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142915665","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 : 2025-02-01DOI: 10.1016/j.vetmic.2024.110362
Anna Ylhäinen , Sari Mölsä , Katariina Thomson , Outi Laitinen-Vapaavuori , Merja Rantala , Thomas Grönthal
Canine pyometra is a common and potentially life-threatening reproductive disorder in intact female dogs. This prospective study aimed to (1) investigate the bacterial spectrum and antimicrobial susceptibilities of bacterial isolates from the uterus and urine of dogs with pyometra, (2) assess the clonal relatedness and virulence factors of Escherichia coli isolates from individual dogs, and (3) determine the occurrence of concurrent and persistent bacteriuria or clinical urinary tract infections. Bacterial isolates from 208 uterine and 203 urine specimens collected during pyometra surgery were analyzed. Additionally, follow-up urine specimens were collected from 56 dogs with perioperative bacteriuria. Bacterial growth was detected in 87 % (180/208) of uterine specimens, while concurrent bacteriuria was observed in 33 % (67/203) of cases. In one-third (18/56) of these dogs the bacteriuria persisted, being primarily (15/18) asymptomatic. E. coli was the most common isolate in both uterine (71 %) and urine (81 %) specimens. Notably, altogether 32 distinct bacterial species were identified, with mixed growth in 15 % of the specimens. The vast majority of isolates were largely susceptible to tested antimicrobials. Identification of bacterial species was performed using MALDI-ToF MS, and antimicrobial susceptibility was assessed by disk diffusion. Whole-genome sequencing of 45 E. coli strains from fifteen dogs indicated high genetic similarities within individual dogs, supporting a clonal relationship. In conclusion, canine uteri with pyometra contained a plethora of bacterial species, predominantly E. coli, and antimicrobial resistance was rare. Concurrent and persistent E. coli bacteriuria was commonly caused by the same clone as found in the uterus.
{"title":"Bacteria associated with canine pyometra and concurrent bacteriuria: A prospective study","authors":"Anna Ylhäinen , Sari Mölsä , Katariina Thomson , Outi Laitinen-Vapaavuori , Merja Rantala , Thomas Grönthal","doi":"10.1016/j.vetmic.2024.110362","DOIUrl":"10.1016/j.vetmic.2024.110362","url":null,"abstract":"<div><div>Canine pyometra is a common and potentially life-threatening reproductive disorder in intact female dogs. This prospective study aimed to (1) investigate the bacterial spectrum and antimicrobial susceptibilities of bacterial isolates from the uterus and urine of dogs with pyometra, (2) assess the clonal relatedness and virulence factors of <em>Escherichia coli</em> isolates from individual dogs, and (3) determine the occurrence of concurrent and persistent bacteriuria or clinical urinary tract infections. Bacterial isolates from 208 uterine and 203 urine specimens collected during pyometra surgery were analyzed. Additionally, follow-up urine specimens were collected from 56 dogs with perioperative bacteriuria. Bacterial growth was detected in 87 % (180/208) of uterine specimens, while concurrent bacteriuria was observed in 33 % (67/203) of cases. In one-third (18/56) of these dogs the bacteriuria persisted, being primarily (15/18) asymptomatic. <em>E. coli</em> was the most common isolate in both uterine (71 %) and urine (81 %) specimens. Notably, altogether 32 distinct bacterial species were identified, with mixed growth in 15 % of the specimens. The vast majority of isolates were largely susceptible to tested antimicrobials. Identification of bacterial species was performed using MALDI-ToF MS, and antimicrobial susceptibility was assessed by disk diffusion. Whole-genome sequencing of 45 <em>E. coli</em> strains from fifteen dogs indicated high genetic similarities within individual dogs, supporting a clonal relationship. In conclusion, canine uteri with pyometra contained a plethora of bacterial species, predominantly <em>E. coli</em>, and antimicrobial resistance was rare. Concurrent and persistent <em>E. coli</em> bacteriuria was commonly caused by the same clone as found in the uterus.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"301 ","pages":"Article 110362"},"PeriodicalIF":2.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142955565","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 : 2025-02-01DOI: 10.1016/j.vetmic.2025.110377
Ibrahim Elsohaby , Polychronis Kostoulas , Mahmoud Fayez , Ahmed Elmoslemany , Mohamed E. Alkafafy , Ahmad M. Bahhary , Reham Alzahrani , Abd El Karem M. Morsi , Juan Carlos Arango-Sabogal
Paratuberculosis, a chronic wasting disease affecting domestic and wild ruminants worldwide, is caused by Mycobacterium avium subsp. paratuberculosis (MAP). Various diagnostic tests exist for detecting MAP infection; however, none of them possess perfect accuracy to be qualified as a reference standard test, particularly due to their notably low sensitivity. Therefore, we used Bayesian latent class models (BLCMs) to estimate diagnostic accuracy of fecal smears (FS), fecal PCR and serum ELISA for detecting MAP infections in sheep, goats, cattle, and camels older than 2 years in Saudi Arabia. Data from a cross-sectional study conducted in the Eastern Province of Saudi Arabia on 31 different farms with a history of MAP infection were analyzed. Fecal and blood samples from all animals older than 2 years in each farm were collected, resulting in a total of 220 sheep, 123 goats, 66 cattle, and 240 camels sampled. FS and IS900-PCR were performed on fecal samples to detect acid-fast bacilli and MAP DNA, respectively. The IDEXX ELISA kit was used to detect MAP antibodies in serum samples. For each ruminant species population, a BLCM was fitted to obtain posterior estimates [medians and 95 % Bayesian credible intervals (95 % BCI)] for sensitivity (Se) and specificity (Sp) of the three tests. We assumed FS and PCR to be conditionally dependent on the true animal MAP status. Prior distributions for test accuracy were used if available. FS had the highest Se among all tests and across all species with median values around 80 % in sheep, goats and camels, and near 50 % in cattle. Median Sp estimates of ELISA and PCR were higher than 90 % for all species. FS yielded the lowest Sp of the study when applied in camels, sheep, and goats. Using the prevalence observed in this study, median positive predictive value (PPV) was higher for PCR and ELISA than FS for camels, sheep, and goats. In cattle, PPV of all tests was similar with median estimates > 95 %. In camels, sheep, and goats, median negative predicative value (NPV) of all tests were > 60 %. The lowest median NPV for all tests were observed in cattle (< 30 %). Our results suggest that ELISA is a suitable option to identify MAP infected animals in farms with previous history of MAP in the Eastern region of Saudi Arabia.
{"title":"Bayesian estimation of diagnostic accuracy of fecal smears, fecal PCR and serum ELISA for detecting Mycobacterium avium subsp. paratuberculosis infections in four domestic ruminant species in Saudi Arabia","authors":"Ibrahim Elsohaby , Polychronis Kostoulas , Mahmoud Fayez , Ahmed Elmoslemany , Mohamed E. Alkafafy , Ahmad M. Bahhary , Reham Alzahrani , Abd El Karem M. Morsi , Juan Carlos Arango-Sabogal","doi":"10.1016/j.vetmic.2025.110377","DOIUrl":"10.1016/j.vetmic.2025.110377","url":null,"abstract":"<div><div>Paratuberculosis, a chronic wasting disease affecting domestic and wild ruminants worldwide, is caused by <em>Mycobacterium avium</em> subsp. <em>paratuberculosis</em> (MAP). Various diagnostic tests exist for detecting MAP infection; however, none of them possess perfect accuracy to be qualified as a reference standard test, particularly due to their notably low sensitivity. Therefore, we used Bayesian latent class models (BLCMs) to estimate diagnostic accuracy of fecal smears (FS), fecal PCR and serum ELISA for detecting MAP infections in sheep, goats, cattle, and camels older than 2 years in Saudi Arabia. Data from a cross-sectional study conducted in the Eastern Province of Saudi Arabia on 31 different farms with a history of MAP infection were analyzed. Fecal and blood samples from all animals older than 2 years in each farm were collected, resulting in a total of 220 sheep, 123 goats, 66 cattle, and 240 camels sampled. FS and IS<em>900</em>-PCR were performed on fecal samples to detect acid-fast bacilli and MAP DNA, respectively. The IDEXX ELISA kit was used to detect MAP antibodies in serum samples. For each ruminant species population, a BLCM was fitted to obtain posterior estimates [medians and 95 % Bayesian credible intervals (95 % BCI)] for sensitivity (Se) and specificity (Sp) of the three tests. We assumed FS and PCR to be conditionally dependent on the true animal MAP status. Prior distributions for test accuracy were used if available. FS had the highest Se among all tests and across all species with median values around 80 % in sheep, goats and camels, and near 50 % in cattle. Median Sp estimates of ELISA and PCR were higher than 90 % for all species. FS yielded the lowest Sp of the study when applied in camels, sheep, and goats. Using the prevalence observed in this study, median positive predictive value (PPV) was higher for PCR and ELISA than FS for camels, sheep, and goats. In cattle, PPV of all tests was similar with median estimates > 95 %. In camels, sheep, and goats, median negative predicative value (NPV) of all tests were > 60 %. The lowest median NPV for all tests were observed in cattle (< 30 %). Our results suggest that ELISA is a suitable option to identify MAP infected animals in farms with previous history of MAP in the Eastern region of Saudi Arabia.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"301 ","pages":"Article 110377"},"PeriodicalIF":2.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142955566","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 : 2025-02-01DOI: 10.1016/j.vetmic.2024.110344
Meire Maria da Silva , Fábio Parra Sellera , João Pedro Rueda Furlan , Valentina Aravena-Ramírez , Danny Fuentes-Castillo , Bruna Fuga , Alexandre José dos Santos Fróes , Alana Lislea de Sousa , Felício Garino Junior , Nilton Lincopan
Extended-spectrum β-lactamase (ESBL)-producing Escherichia coli poses a significant threat to public health due to its ability to confer resistance to broad-spectrum antimicrobials, especially third-generation cephalosporins. Herein, we report gut colonization by international clones of CTX-M-8-producing E. coli in scorpion mud turtles (Kinosternon scorpioides) from a captive breeding program in the Brazilian Amazon. The E. coli strains exhibited multidrug resistance to clinically relevant antimicrobials, and genomic analyses revealed broad resistomes to antimicrobials, heavy metals, pesticides, and disinfectants. Detecting these medically important bacteria in captive wildlife underscores the potential risks associated with reintroduction programs, as ESBL-producing bacteria may spill over into the natural ecosystem and threaten wildlife. Our findings highlight the need for enhanced surveillance and control measures in wildlife conservation programs to mitigate the risks of antimicrobial resistance transmission between captive and wild populations.
产广谱β-内酰胺酶（ESBL）的大肠杆菌能够对广谱抗菌素，特别是第三代头孢菌素产生耐药性，对公众健康构成重大威胁。在此，我们报告了产ctx - m -8大肠杆菌的国际克隆在巴西亚马逊圈养繁殖计划的蝎泥龟（Kinosternon scorpioides）的肠道定植。大肠杆菌菌株对临床相关的抗菌素表现出多重耐药性，基因组分析显示对抗菌素、重金属、杀虫剂和消毒剂具有广泛的抗性。在圈养野生动物身上检测到这些医学上重要的细菌，强调了与放归计划相关的潜在风险，因为产生esbl的细菌可能会溢出到自然生态系统中，威胁野生动物。我们的研究结果强调了在野生动物保护计划中加强监测和控制措施的必要性，以减轻圈养种群和野生种群之间抗菌素耐药性传播的风险。
{"title":"Gut colonization of semi-aquatic turtles inhabiting the Brazilian Amazon by international clones of CTX-M-8-producing Escherichia coli","authors":"Meire Maria da Silva , Fábio Parra Sellera , João Pedro Rueda Furlan , Valentina Aravena-Ramírez , Danny Fuentes-Castillo , Bruna Fuga , Alexandre José dos Santos Fróes , Alana Lislea de Sousa , Felício Garino Junior , Nilton Lincopan","doi":"10.1016/j.vetmic.2024.110344","DOIUrl":"10.1016/j.vetmic.2024.110344","url":null,"abstract":"<div><div>Extended-spectrum β-lactamase (ESBL)-producing <em>Escherichia coli</em> poses a significant threat to public health due to its ability to confer resistance to broad-spectrum antimicrobials, especially third-generation cephalosporins. Herein, we report gut colonization by international clones of CTX-M-8-producing <em>E. coli</em> in scorpion mud turtles (<em>Kinosternon scorpioides</em>) from a captive breeding program in the Brazilian Amazon. The <em>E. coli</em> strains exhibited multidrug resistance to clinically relevant antimicrobials, and genomic analyses revealed broad resistomes to antimicrobials, heavy metals, pesticides, and disinfectants. Detecting these medically important bacteria in captive wildlife underscores the potential risks associated with reintroduction programs, as ESBL-producing bacteria may spill over into the natural ecosystem and threaten wildlife. Our findings highlight the need for enhanced surveillance and control measures in wildlife conservation programs to mitigate the risks of antimicrobial resistance transmission between captive and wild populations.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"301 ","pages":"Article 110344"},"PeriodicalIF":2.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142886159","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 : 2025-02-01DOI: 10.1016/j.vetmic.2024.110360
Peng Gao , Jianle Ren , Qiongqiong Zhou, Peng Chen, Ailin Zhang, Yongning Zhang, Lei Zhou, Xinna Ge, Xin Guo, Jun Han, Hanchun Yang
Pseudorabies virus (PRV) poses a significant threat to the global swine breeding industry and public health, but how the virus transverses the host defense systems for efficient viral replication and pathogenesis remains unclear. Here, we report that PRV could inhibit the unfolded protein response (UPR), a critical component of host innate immunity against viral infection, to promote virus replication during the late infection stages. PERK was shown phosphorylated and active in PRV-infected cells, but the subsequent events were suppressed post virus infection, such as eIF2α phosphorylation, ATF4 expression, and the formation of stress granules (SGs). In the meantime, although IRE1α was also active, its activated effector XBP1s was suppressed through downregulation of XBP1 mRNA levels and cleavage of XBP1s protein. Our findings also indicate that the Golgi apparatus, where ATF6 activation occur, was severely damaged in PRV-infected cells. Meanwhile, the downstream regulatory genes associated with the three UPR sensors, such as ERp60, CHOP, and EDEM1, remained silent in PRV-infected cells. Enhanced viral replication was observed post knockdown of UPR effectors ATF4 or XBP1, while stimulation with UPR activators inhibits virus replication. In conclusion, our findings address the critical question of how PRV regulates cellular UPR in favor of viral replication, and expand understanding of viruses mediated UPR suppression in general.
{"title":"Pseudorabies virus inhibits the unfolded protein response for viral replication during the late stages of infection","authors":"Peng Gao , Jianle Ren , Qiongqiong Zhou, Peng Chen, Ailin Zhang, Yongning Zhang, Lei Zhou, Xinna Ge, Xin Guo, Jun Han, Hanchun Yang","doi":"10.1016/j.vetmic.2024.110360","DOIUrl":"10.1016/j.vetmic.2024.110360","url":null,"abstract":"<div><div>Pseudorabies virus (PRV) poses a significant threat to the global swine breeding industry and public health, but how the virus transverses the host defense systems for efficient viral replication and pathogenesis remains unclear. Here, we report that PRV could inhibit the unfolded protein response (UPR), a critical component of host innate immunity against viral infection, to promote virus replication during the late infection stages. PERK was shown phosphorylated and active in PRV-infected cells, but the subsequent events were suppressed post virus infection, such as eIF2α phosphorylation, ATF4 expression, and the formation of stress granules (SGs). In the meantime, although IRE1α was also active, its activated effector XBP1s was suppressed through downregulation of XBP1 mRNA levels and cleavage of XBP1s protein. Our findings also indicate that the Golgi apparatus, where ATF6 activation occur, was severely damaged in PRV-infected cells. Meanwhile, the downstream regulatory genes associated with the three UPR sensors, such as ERp60, CHOP, and EDEM1, remained silent in PRV-infected cells. Enhanced viral replication was observed post knockdown of UPR effectors ATF4 or XBP1, while stimulation with UPR activators inhibits virus replication. In conclusion, our findings address the critical question of how PRV regulates cellular UPR in favor of viral replication, and expand understanding of viruses mediated UPR suppression in general.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"301 ","pages":"Article 110360"},"PeriodicalIF":2.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142932673","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 : 2025-02-01DOI: 10.1016/j.vetmic.2024.110363
Paulina Śliwka , David Sáez Moreno , Paweł Korzeniowski , Agata Milcarz , Maciej Kuczkowski , Rafał Kolenda , Sylwia Kozioł , Magdalena Narajczyk , Uwe Roesler , Ludwika Tomaszewska-Hetman , Marta Kuźmińska-Bajor
Avian pathogenic Escherichia coli (APEC) is a principal etiologic agent of avian colibacillosis, responsible for significant economic losses in the poultry industry due to high mortality and disease treatment with antibiotics. APEC and its ability to form biofilms on food and processing surfaces contributes to its persistence within farms. Bacteriophages are promising antibacterial agents for combating APEC. This study focused on characterization of the newly isolated phages UPWr_E1, UPWr_E2, and UPWr_E4 as well as the UPWr_E124 phage cocktail containing these three phages. Methods included efficiency of plating assay, transmission electron microscopy, and characterization of their resistance to different pH values and temperatures. Moreover, phage genomes were sequenced, annotated and analyzed, and were compared with previously sequenced E. coli phages. All three phages are virulent and devoid of undesirable genes for therapy. Phage UPWr_E1 belongs to the genus Krischvirus within the order Straboviridae and both UPWr_E2 and UPWr_E4 belong to the genus Tequatrovirus within the subfamily Tevenvirinae, sharing over 95 % nucleotide identity between them. For their use on poultry farms, UPWr_E phages and the UPWr_E124 phage cocktail were tested for their anti-biofilm activity on two E. coli strains – 158B (APEC) and the strong biofilm producer NCTC 17848 – on two abiotic surfaces: a 96-well microplate, a stainless steel surface, and one biotic surface, represented by lettuce leaves. The reduction of biofilm formed by both strains in the 96-well microplate, on the stainless steel and lettuce leaf surface for bacteriophage treatment was very efficient, reducing biofilms by ranges of 50.2–83.6, 58.2–88.4 and 53–99.4 %, respectively. Therefore, we conclude that UPWr_E phages and the UPWr_E124 phage cocktail are promising candidates for APEC biocontrol.
{"title":"Avian pathogenic Escherichia coli-targeting phages for biofilm biocontrol in the poultry industry","authors":"Paulina Śliwka , David Sáez Moreno , Paweł Korzeniowski , Agata Milcarz , Maciej Kuczkowski , Rafał Kolenda , Sylwia Kozioł , Magdalena Narajczyk , Uwe Roesler , Ludwika Tomaszewska-Hetman , Marta Kuźmińska-Bajor","doi":"10.1016/j.vetmic.2024.110363","DOIUrl":"10.1016/j.vetmic.2024.110363","url":null,"abstract":"<div><div>Avian pathogenic <em>Escherichia coli</em> (APEC) is a principal etiologic agent of avian colibacillosis, responsible for significant economic losses in the poultry industry due to high mortality and disease treatment with antibiotics. APEC and its ability to form biofilms on food and processing surfaces contributes to its persistence within farms. Bacteriophages are promising antibacterial agents for combating APEC. This study focused on characterization of the newly isolated phages UPWr_E1, UPWr_E2, and UPWr_E4 as well as the UPWr_E124 phage cocktail containing these three phages. Methods included efficiency of plating assay, transmission electron microscopy, and characterization of their resistance to different pH values and temperatures. Moreover, phage genomes were sequenced, annotated and analyzed, and were compared with previously sequenced <em>E. coli</em> phages. All three phages are virulent and devoid of undesirable genes for therapy. Phage UPWr_E1 belongs to the genus <em>Krischvirus</em> within the order <em>Straboviridae</em> and both UPWr_E2 and UPWr_E4 belong to the genus <em>Tequatrovirus</em> within the subfamily <em>Tevenvirinae</em>, sharing over 95 % nucleotide identity between them. For their use on poultry farms, UPWr_E phages and the UPWr_E124 phage cocktail were tested for their anti-biofilm activity on two <em>E. coli</em> strains – 158B (APEC) and the strong biofilm producer NCTC 17848 – on two abiotic surfaces: a 96-well microplate, a stainless steel surface, and one biotic surface, represented by lettuce leaves. The reduction of biofilm formed by both strains in the 96-well microplate, on the stainless steel and lettuce leaf surface for bacteriophage treatment was very efficient, reducing biofilms by ranges of 50.2–83.6, 58.2–88.4 and 53–99.4 %, respectively. Therefore, we conclude that UPWr_E phages and the UPWr_E124 phage cocktail are promising candidates for APEC biocontrol.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"301 ","pages":"Article 110363"},"PeriodicalIF":2.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142967091","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 : 2025-02-01DOI: 10.1016/j.vetmic.2025.110373
Chongyang Wang , Ting Wang , Qingrun He , Qili Hou , Liuyuan Duan , Ruochen Hu , Yu Han , Yongchun Yang , Houhui Song , Zengqi Yang
Pseudorabies virus (PRV) is one of the highly contagious pathogens causing significant economic losses to the swine industry worldwide. More importantly, PRV is becoming a potential “life-threatening zoonosis” since the human-originated PRV strain was first isolated in 2019. Previously we found that the canonical Wnt/β-catenin pathway facilitates PRV proliferation, while the underlying mechanism remains unknown. In this study, the antiviral activities of the Wnt inhibitors (Adavivint, CCT251545, FH535, and iCRT14) were identified. Applying these inhibitors significantly inhibited PRV proliferation in different cell lines. Among them, CCT251545 presented the strongest anti-PRV activity with IC50 values less than 200 nM. Our in vivo studies showed that treatment with CCT251545 remarkedly decreased the viral loads and protected mice challenged with PRV. Further study found that CCT251545 neither had a virucidal effect nor affected viral adsorption while mainly interfering with the entry process of the PRV life cycle. Using the FITC-dextran uptake assay, we found that CCT251545 inhibited macropinocytosis. The formation of membrane protrusions, which is important for macropinocytosis, was also inhibited by CCT251545. Consistent with this, knockout of β-catenin suppressed the PRV macropinocytosis and the formation of protrusions. On the contrary, LiCl treatment significantly stimulated the protrusion formation and the PRV entry. Together, these findings suggest that suppression of the Wnt/β-catenin pathway inhibits the macropinocytosis-dependent entry of PRV, thereby providing potential targets for developing antiviral agents against PRV.
{"title":"Inhibition of the canonical Wnt/β-catenin pathway interferes with macropinocytosis to suppress pseudorabies virus proliferation","authors":"Chongyang Wang , Ting Wang , Qingrun He , Qili Hou , Liuyuan Duan , Ruochen Hu , Yu Han , Yongchun Yang , Houhui Song , Zengqi Yang","doi":"10.1016/j.vetmic.2025.110373","DOIUrl":"10.1016/j.vetmic.2025.110373","url":null,"abstract":"<div><div>Pseudorabies virus (PRV) is one of the highly contagious pathogens causing significant economic losses to the swine industry worldwide. More importantly, PRV is becoming a potential “life-threatening zoonosis” since the human-originated PRV strain was first isolated in 2019. Previously we found that the canonical Wnt/β-catenin pathway facilitates PRV proliferation, while the underlying mechanism remains unknown. In this study, the antiviral activities of the Wnt inhibitors (Adavivint, CCT251545, FH535, and iCRT14) were identified. Applying these inhibitors significantly inhibited PRV proliferation in different cell lines. Among them, CCT251545 presented the strongest anti-PRV activity with IC<sub>50</sub> values less than 200 nM. Our <em>in vivo</em> studies showed that treatment with CCT251545 remarkedly decreased the viral loads and protected mice challenged with PRV. Further study found that CCT251545 neither had a virucidal effect nor affected viral adsorption while mainly interfering with the entry process of the PRV life cycle. Using the FITC-dextran uptake assay, we found that CCT251545 inhibited macropinocytosis. The formation of membrane protrusions, which is important for macropinocytosis, was also inhibited by CCT251545. Consistent with this, knockout of β-catenin suppressed the PRV macropinocytosis and the formation of protrusions. On the contrary, LiCl treatment significantly stimulated the protrusion formation and the PRV entry. Together, these findings suggest that suppression of the Wnt/β-catenin pathway inhibits the macropinocytosis-dependent entry of PRV, thereby providing potential targets for developing antiviral agents against PRV.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"301 ","pages":"Article 110373"},"PeriodicalIF":2.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142967092","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 : 2025-02-01DOI: 10.1016/j.vetmic.2024.110354
Jun Kwon , Sang Guen Kim , Sang Wha Kim , Hyoun Joong Kim , Jung Woo Kang , Su Jin Jo , Sib Sankar Giri , Won Jun Jeong , Sung Bin Lee , Ji Hyung Kim , Se Chang Park
Canine otitis externa, characterized by the involvement of diverse bacterial species, notably Pseudomonas aeruginosa and Staphylococcus pseudintermedius, necessitates antibiotic administration as the primary therapeutic approach; however, prolonged treatment often precipitates antibiotic resistance. Therefore, the application of bacteriophages as antimicrobial agents has been of interest recently. However, phage therapy has limitations; its efficacy depends on the lytic capacity of the phage and the emergence of phage resistance, which can be overcome by using phage cocktails. This study aimed to enhance the therapeutic potential of bacteriophages by supplementing additional materials to hinder the pathogens and combining different viruses to broaden the lytic spectrum. The therapeutic potential of the phage cocktail, consisting of Pseudomonas phage pPa_SNUABM_DT01 and Staphylococcus phage pSp_SNUABM-J, was evaluated using an in vitro planktonic bacterial cell lysis assay and a biofilm degradation assay. Additionally, its efficacy was assessed using an in vivo mouse otitis externa model and clinical administration in five dogs with chronic Pseudomonas aeruginosa and Staphylococcus pseudintermedius otitis externa. The phage cocktail with formulation, including glycerol, glycine, and Tween 20, as additional components resulted in a significant reduction in bacterial counts and clinical improvements, including odor, discharge type and amount, and inflammatory symptoms. The results suggest that administering a phage cocktail solution with additional components could make phage therapy a more efficient treatment for otitis externa in dogs. This offers a practical alternative to traditional antibiotic treatments and could help mitigate antibiotic resistance in veterinary medicine.
{"title":"Tailoring formulation for enhanced phage therapy in canine otitis externa: a cocktail approach targeting Pseudomonas aeruginosa and Staphylococcus pseudintermedius","authors":"Jun Kwon , Sang Guen Kim , Sang Wha Kim , Hyoun Joong Kim , Jung Woo Kang , Su Jin Jo , Sib Sankar Giri , Won Jun Jeong , Sung Bin Lee , Ji Hyung Kim , Se Chang Park","doi":"10.1016/j.vetmic.2024.110354","DOIUrl":"10.1016/j.vetmic.2024.110354","url":null,"abstract":"<div><div>Canine otitis externa, characterized by the involvement of diverse bacterial species, notably <em>Pseudomonas aeruginosa</em> and <em>Staphylococcus pseudintermedius</em>, necessitates antibiotic administration as the primary therapeutic approach; however, prolonged treatment often precipitates antibiotic resistance. Therefore, the application of bacteriophages as antimicrobial agents has been of interest recently. However, phage therapy has limitations; its efficacy depends on the lytic capacity of the phage and the emergence of phage resistance, which can be overcome by using phage cocktails. This study aimed to enhance the therapeutic potential of bacteriophages by supplementing additional materials to hinder the pathogens and combining different viruses to broaden the lytic spectrum. The therapeutic potential of the phage cocktail, consisting of <em>Pseudomonas</em> phage pPa_SNUABM_DT01 and <em>Staphylococcus</em> phage pSp_SNUABM-J, was evaluated using an in vitro planktonic bacterial cell lysis assay and a biofilm degradation assay. Additionally, its efficacy was assessed using an <em>in vivo</em> mouse otitis externa model and clinical administration in five dogs with chronic <em>Pseudomonas aeruginosa</em> and <em>Staphylococcus pseudintermedius</em> otitis externa. The phage cocktail with formulation, including glycerol, glycine, and Tween 20, as additional components resulted in a significant reduction in bacterial counts and clinical improvements, including odor, discharge type and amount, and inflammatory symptoms. The results suggest that administering a phage cocktail solution with additional components could make phage therapy a more efficient treatment for otitis externa in dogs. This offers a practical alternative to traditional antibiotic treatments and could help mitigate antibiotic resistance in veterinary medicine.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"301 ","pages":"Article 110354"},"PeriodicalIF":2.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142910933","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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