Objective: This study assessed the applicability of loop-mediated isothermal amplification (LAMP) for the detection of leptospirosis among domesticated animals and sewage rats. Specifically, it evaluated the ability of LAMP to amplify Leptospira spp. targeting the 16s rRNA gene in boiled urine samples. Materials and methods: A total of 140 samples from different domestic animals were tested for the presence of the antigen. A nested-polymerase chain reaction (nPCR) protocol was used to compare and determine the sensitivity of LAMP in detecting Leptospira spp. The LAMP was also evaluated by comparing its amplification result using agarose gel electrophoresis and color change using dye. Results: Positivity rate of Leptospira spp. antigen was 29.0% (40/140) for LAMP and 9.3% (13/140) for nPCR. Also, LAMP results for gel electrophoresis and dye color change varied in some samples that may be due to the interpretation of the result in dye color change. Conclusion: Overall, LAMP is a rapid, sensitive, and cost-effective diagnostic method compared with nPCR. Also, LAMP has a potential application as pen-side screening, surveillance, and clinical diagnostic kits of infectious diseases without requiring advance equipment and skilled personnel. http://doi.org/10.5455/javar.2018.e299

Objective: This study assessed the applicability of loop-mediated isothermal amplification (LAMP) for the detection of leptospirosis among domesticated animals and sewage rats. Specifically, it evaluated the ability of LAMP to amplify Leptospira spp. targeting the 16s rRNA gene in boiled urine samples. Materials and methods: A total of 140 samples from different domestic animals were tested for the presence of the antigen. A nested-polymerase chain reaction (nPCR) protocol was used to compare and determine the sensitivity of LAMP in detecting Leptospira spp. The LAMP was also evaluated by comparing its amplification result using agarose gel electrophoresis and color change using dye. Results: Positivity rate of Leptospira spp. antigen was 29.0% (40/140) for LAMP and 9.3% (13/140) for nPCR. Also, LAMP results for gel electrophoresis and dye color change varied in some samples that may be due to the interpretation of the result in dye color change. Conclusion: Overall, LAMP is a rapid, sensitive, and cost-effective diagnostic method compared with nPCR. Also, LAMP has a potential application as pen-side screening, surveillance, and clinical diagnostic kits of infectious diseases without requiring advance equipment and skilled personnel. [J Adv Vet Anim Res 2018; 5(4.000): 459-465]

Objective: The aim of this paper is to report surgical correction of ureteric rupture due to stenosis induced secondary to accidental injury by placing nephrovesical subcutaneous ureteric bypass in a dog and postoperative long term outcomes.Materials and methods: Imaging techniques revealed multiple bone fractures as well as left proximal ureter injury. The all bone fractures were corrected using standard techniques while left proximal ureter injury was treated as conservation medical therapy. One month later, contrast media were also found in proximal ureter and abdomen which indicated ureter rupture. This rupture was corrected surgically by nephrovesical subcutaneous ureteric bypass (SUB) under fluoroscopy guidance.Results: First day after accidental injury, the serum BUN and CRE were 10.7 mg/dL and 0.9 mg/dL, respectively which indicated kidney injury but by conservative therapy these parameters were lowered gradually. On the 5th day after considering these parameters, the dog was judged normal. However, on the 31st day BUN and CRE were 14.3 mg/dL and 0.8 mg/Kg, respectively. The Doppler ultrasonography revealed hydroneprosis, proximal ureter stenosis and high resistive index (0.72±0.02) in the renal arcuate artery indicating renal abnormalities due to ureter obstruction. On re-examination by radiography after one month postoperatively, revealed that patency of the device and normal renal function. The dog was clinically normal with normal urination and no complications were found 6 months postoperatively.Conclusion: In view of the above findings, it is suggested that the SUB system can be a better alternative to preserve the kidney in non-reparable traumatic ureteral damage in dogs.http://doi.org/10.5455/javar.2018.e266

Objective: The aim of this paper is to report surgical correction of ureteric rupture due to stenosis induced secondary to accidental injury by placing nephrovesical subcutaneous ureteric bypass in a dog and postoperative long term outcomes. Materials and methods: Imaging techniques revealed multiple bone fractures as well as left proximal ureter injury. The all bone fractures were corrected using standard techniques while left proximal ureter injury was treated as conservation medical therapy. One month later, contrast media were also found in proximal ureter and abdomen which indicated ureter rupture. This rupture was corrected surgically by nephrovesical subcutaneous ureteric bypass (SUB) under fluoroscopy guidance. Results: First day after accidental injury, the serum BUN and CRE were 10.7 mg/dL and 0.9 mg/dL, respectively which indicated kidney injury but by conservative therapy these parameters were lowered gradually. On the 5th day after considering these parameters, the dog was judged normal. However, on the 31st day BUN and CRE were 14.3 mg/dL and 0.8 mg/Kg, respectively. The Doppler ultrasonography revealed hydroneprosis, proximal ureter stenosis and high resistive index (0.72±0.02) in the renal arcuate artery indicating renal abnormalities due to ureter obstruction. On re-examination by radiography after one month postoperatively, revealed that patency of the device and normal renal function. The dog was clinically normal with normal urination and no complications were found 6 months postoperatively. Conclusion: In view of the above findings, it is suggested that the SUB system can be a better alternative to preserve the kidney in non-reparable traumatic ureteral damage in dogs. [J Adv Vet Anim Res 2018; 5(2.000): 247-254]

Objective: The main aim of this case report is to present a case of feline hemotropic mycoplasmosis that occurred concurrently with bacterial cystitis following the bite. Material and methods: A 3-year-old intact male domestic shorthair cat weighing 3.7 kg was referred to the Universiti Malaysia Kelantan Veterinary Clinic with clinical signs of hematuria and dysuria. History revealed that it was managed outdoor, fed with kibbles and wet food, but with no vaccination and deworming. Upon physical examination, the cat had a dull appearance, pale mucous membrane, normal respiratory rate, hypothermia, and bradycardia. Upon the examination of the urogenital system, there were urine burns at the anal region, necrotized penile tip, and presence of bite wound observed at the perineal region. Turgid and enlarged urinary bladder was identified upon palpation. Results: Diagnostic investigation revealed the hemotropic mycoplasmosis via microscopy, while urine culture was positive for Escherichia coli infection. The cat was successfully treated symptomatically. Conclusion: However, the prognosis of this cat was guarded given that the anemia was unresolved at the point of discharge. http://doi.org/10.5455/javar.2018.e304

Objective: The main aim of this case report is to present a case of feline hemotropic mycoplasmosis that occurred concurrently with bacterial cystitis following the bite. Material and methods: A 3-year-old intact male domestic shorthair cat weighing 3.7 kg was referred to the Universiti Malaysia Kelantan Veterinary Clinic with clinical signs of hematuria and dysuria. History revealed that it was managed outdoor, fed with kibbles and wet food, but with no vaccination and deworming. Upon physical examination, the cat had a dull appearance, pale mucous membrane, normal respiratory rate, hypothermia, and bradycardia. Upon the examination of the urogenital system, there were urine burns at the anal region, necrotized penile tip, and presence of bite wound observed at the perineal region. Turgid and enlarged urinary bladder was identified upon palpation. Results: Diagnostic investigation revealed the hemotropic mycoplasmosis via microscopy, while urine culture was positive for Escherichia coli infection. The cat was successfully treated symptomatically. Conclusion: However, the prognosis of this cat was guarded given that the anemia was unresolved at the point of discharge. [J Adv Vet Anim Res 2018; 5(4.000): 490-495]

Enterococcus species have developed from being commensal bacteria to leading pathogens that cause infections in humans and animals. The gastrointestinal tract of mammals is the normal habitat of these species. Virulence factors are proteins that are produced by the bacterium which are used to enhance their pathogenicity. The objectives of this study were to isolate Enterococcus spp. from livestock and companion animals, differentiate between the different sub-species and detect the presence of important virulence genes. Rectal and saliva swabs were collected from dogs and cats, whereas only rectal swabs were collected from cattle and cloacal swabs from chickens. Presumptive Enterococcus was selected using Bile Esculin Azide (BEA) agar, and Enterococcus species were confirmed using the polymerase chain reaction (PCR) by amplifying the tuf gene. In order to differentiate between E. faecalis and E. faecium, a multiplex PCR was used to detect the SodA gene. The genes responsible for gelatinase production (gelE) and for conjugation (ccf) were also detected using PCR. Out of 211 animal swabs, 182 (86%) were positive for the tuf gene. Overall, there were 55 isolates of E. faecalis (30%) compared to 22 isolates of E. faecium (12%). The virulence genes had a prevalence of 52% and 36% for gelE and ccf, respectively, in all animal hosts. The results demonstrated that chicken cloacal samples had the highest prevalence for E. faecalis, gelE and ccf genes compared to all the other isolates detected from other animal hosts. The results also demonstrated a statistically significant (p  0.05) association between the prevalence of virulence genes (gelE and ccf) and animal species from which Enterococcus spp. was isolated. We provided evidence that healthy livestock and companion animals can harbour pathogenic Enterococcus that can be transferred via the food chain as well as through close association such as petting and licking of humans. This study partially demonstrated that Enterococci spp. are capable of evolving from being simple commensal bacteria to becoming pathogens that cause infection in humans and animals through the acquisition of virulence factors through mobile genetic elements.

Enterococcus species have developed from being commensal bacteria to leading pathogens that cause infections in humans and animals. The gastrointestinal tract of mammals is the normal habitat of these species. Virulence factors are proteins that are produced by the bacterium which are used to enhance their pathogenicity. The objectives of this study were to isolate Enterococcus spp. from livestock and companion animals, differentiate between the different sub-species and detect the presence of important virulence genes. Rectal and saliva swabs were collected from dogs and cats, whereas only rectal swabs were collected from cattle and cloacal swabs from chickens. Presumptive Enterococcus was selected using Bile Esculin Azide (BEA) agar, and Enterococcus species were confirmed using the polymerase chain reaction (PCR) by amplifying the tuf gene. In order to differentiate between E. faecalis and E. faecium, a multiplex PCR was used to detect the SodA gene. The genes responsible for gelatinase production (gelE) and for conjugation (ccf) were also detected using PCR. Out of 211 animal swabs, 182 (86%) were positive for the tuf gene. Overall, there were 55 isolates of E. faecalis (30%) compared to 22 isolates of E. faecium (12%). The virulence genes had a prevalence of 52% and 36% for gelE and ccf, respectively, in all animal hosts. The results demonstrated that chicken cloacal samples had the highest prevalence for E. faecalis, gelE and ccf genes compared to all the other isolates detected from other animal hosts. The results also demonstrated a statistically significant (p < 0.05) association between the prevalence of virulence genes (gelE and ccf) and animal species from which Enterococcus spp. was isolated. We provided evidence that healthy livestock and companion animals can harbour pathogenic Enterococcus that can be transferred via the food chain as well as through close association such as petting and licking of humans. This study partially demonstrated that Enterococci spp. are capable of evolving from being simple commensal bacteria to becoming pathogens that cause infection in humans and animals through the acquisition of virulence factors through mobile genetic elements.

Virulence-associated genes have been recognised and detected in Campylobacter species. The majority of them have been proven to be associated with pathogenicity. This study aimed to detect the presence of virulence genes associated with pathogenicity and responsible for invasion, expression of adherence, colonisation and production of the cytolethal distending toxin (cdt) in Campylobacter jejuni and Campylobacter coli. Commercial chicken faecal samples were randomly sampled from chicken farms within the Durban metropolitan area in South Africa. Furthermore, human clinical Campylobacter spp. isolates were randomly sampled from a private pathology laboratory in South Africa. Out of a total of 100 chicken faecal samples, 78% (n = 78) were positive for Campylobacter growth on modified charcoal cefoperazone deoxycholate and from the random laboratory collection of 100 human clinical isolates, 83% (n = 83) demonstrated positive Campylobacter spp. growth following culturing methods. These samples were screened for the presence of the following virulence genes: cadF, hipO, asp, ciaB, dnaJ, pldA, cdtA, cdtB and cdtC. As expected, the cadF gene was present in 100% of poultry (n = 78) and human clinical isolates (n = 83). Campylobacter jejuni was the main species detected in both poultry and human clinical isolates, whilst C. coli were detected at a significantly lower percentage (p 0.05). Eight per cent of the C. jejuni from human clinical isolates had all virulence genes that were investigated. Only one C. coli isolate demonstrated the presence of all the virulence genes investigated; however, the pldA virulence gene was detected in 100% of the C. coli isolates in poultry and a high percentage (71%) in human clinical C. coli isolates as well. The detection of cdt genes was found at higher frequency in poultry than human clinical isolates. The high prevalence rates of virulence genes detected in poultry and human clinical isolates demonstrate their significance in the pathogenicity of Campylobacter species.

Virulence-associated genes have been recognised and detected in Campylobacter species. The majority of them have been proven to be associated with pathogenicity. This study aimed to detect the presence of virulence genes associated with pathogenicity and responsible for invasion, expression of adherence, colonisation and production of the cytolethal distending toxin (cdt) in Campylobacter jejuni and Campylobacter coli. Commercial chicken faecal samples were randomly sampled from chicken farms within the Durban metropolitan area in South Africa. Furthermore, human clinical Campylobacter spp. isolates were randomly sampled from a private pathology laboratory in South Africa. Out of a total of 100 chicken faecal samples, 78% (n = 78) were positive for Campylobacter growth on modified charcoal cefoperazone deoxycholate and from the random laboratory collection of 100 human clinical isolates, 83% (n = 83) demonstrated positive Campylobacter spp. growth following culturing methods. These samples were screened for the presence of the following virulence genes: cadF, hipO, asp, ciaB, dnaJ, pldA, cdtA, cdtB and cdtC. As expected, the cadF gene was present in 100% of poultry (n = 78) and human clinical isolates (n = 83). Campylobacter jejuni was the main species detected in both poultry and human clinical isolates, whilst C. coli were detected at a significantly lower percentage (p < 0.05). Eight per cent of the C. jejuni from human clinical isolates had all virulence genes that were investigated. Only one C. coli isolate demonstrated the presence of all the virulence genes investigated; however, the pldA virulence gene was detected in 100% of the C. coli isolates in poultry and a high percentage (71%) in human clinical C. coli isolates as well. The detection of cdt genes was found at higher frequency in poultry than human clinical isolates. The high prevalence rates of virulence genes detected in poultry and human clinical isolates demonstrate their significance in the pathogenicity of Campylobacter species.

Lactococcus garvieae is a Gram-positive bacterium that causes mortalities in freshwater and marine fish worldwide and therefore results in severe economic losses in the aquaculture industry. Apart from the apparent integral role of the exopolysaccharide (EPS) capsule in pathogenesis, factors associated with virulence of this bacterium are poorly understood. However, recent studies have indicated that the ability of L. garvieae to cause disease does not depend on the presence of the EPS capsule. Lack of knowledge of virulence factors, pathogenesis and serology of L. garvieae is an impediment to the development of effective typing methods and control measures. This study, therefore, aimed to detect the presence of EPS capsules and other putative virulence factors in South African L. garvieae fish pathogenic isolates and a non-virulent isolate, and to identify possible candidates for subunit vaccine development. No indication of the presence of the EPS capsule was detected by negative staining or amplification of the EPS biosynthesis gene cluster in the virulent isolates or the avirulent strain, discrediting the notion that the EPS capsule is the sole determinant of virulence. However, a set of putative virulence factor genes was detected in all isolates, and candidates for subunit vaccine development (enolase, lactate dehydrogenase phosphoenolpyruvate-protein phosphotransferase) were identified by identification of extracellular proteins of virulent strains.

Lactococcus garvieae is a Gram-positive bacterium that causes mortalities in freshwater and marine fish worldwide and therefore results in severe economic losses in the aquaculture industry. Apart from the apparent integral role of the exopolysaccharide (EPS) capsule in pathogenesis, factors associated with virulence of this bacterium are poorly understood. However, recent studies have indicated that the ability of L. garvieae to cause disease does not depend on the presence of the EPS capsule. Lack of knowledge of virulence factors, pathogenesis and serology of L. garvieae is an impediment to the development of effective typing methods and control measures. This study, therefore, aimed to detect the presence of EPS capsules and other putative virulence factors in South African L. garvieae fish pathogenic isolates and a non-virulent isolate, and to identify possible candidates for subunit vaccine development. No indication of the presence of the EPS capsule was detected by negative staining or amplification of the EPS biosynthesis gene cluster in the virulent isolates or the avirulent strain, discrediting the notion that the EPS capsule is the sole determinant of virulence. However, a set of putative virulence factor genes was detected in all isolates, and candidates for subunit vaccine development (enolase, lactate dehydrogenase phosphoenolpyruvate-protein phosphotransferase) were identified by identification of extracellular proteins of virulent strains.

Anthrax, a neglected zoonotic disease that is transmitted by a spore-forming, rod-shaped bacterium, Bacillus anthracis, has reached endemic proportions in the Western Province of Zambia. Transmission of anthrax from the environment as well as between cattle has been observed to be partly because of entrenched beliefs, perceptions and traditional practices among cattle farmers in the known outbreak areas. This study was aimed at exploring lay perceptions, beliefs and practices that influence anthrax transmission in cattle of the Western Province. A mixed-methods study was conducted from August to December 2015. Quantitative data were collected using a cross-sectional survey. Qualitative data were generated by interviewing professional staff and community members. Five focus group discussions and five key informant interviews were conducted. Thematic analysis of interview data was performed using NVivo software. The findings suggested that cattle anthrax was biologically as well as culturally maintained. Cattle farmers were reluctant to have their livestock vaccinated against anthrax because of perceived low efficacy of the vaccine. Also, the cattle farmers did not trust professional staff and their technical interventions. Popular cultural practices that involved exchange of animals between herds contributed to uncontrolled cattle movements between herds and subsequent transmission of anthrax. These findings imply the need for professional staff to be culturally competent in handling socio-cultural issues that are known to be barriers for disease control in animals. There is a need to develop a policy framework that will foster integrated control of anthrax across disciplines.

Anthrax, a neglected zoonotic disease that is transmitted by a spore-forming, rod-shaped bacterium, Bacillus anthracis, has reached endemic proportions in the Western Province of Zambia. Transmission of anthrax from the environment as well as between cattle has been observed to be partly because of entrenched beliefs, perceptions and traditional practices among cattle farmers in the known outbreak areas. This study was aimed at exploring lay perceptions, beliefs and practices that influence anthrax transmission in cattle of the Western Province. A mixed-methods study was conducted from August to December 2015. Quantitative data were collected using a cross-sectional survey. Qualitative data were generated by interviewing professional staff and community members. Five focus group discussions and five key informant interviews were conducted. Thematic analysis of interview data was performed using NVivo software. The findings suggested that cattle anthrax was biologically as well as culturally maintained. Cattle farmers were reluctant to have their livestock vaccinated against anthrax because of perceived low efficacy of the vaccine. Also, the cattle farmers did not trust professional staff and their technical interventions. Popular cultural practices that involved exchange of animals between herds contributed to uncontrolled cattle movements between herds and subsequent transmission of anthrax. These findings imply the need for professional staff to be culturally competent in handling socio-cultural issues that are known to be barriers for disease control in animals. There is a need to develop a policy framework that will foster integrated control of anthrax across disciplines.

Avian paramyxovirus type-1 (APMV-1) viruses of the lentogenic pathotypes are often isolated from wild aquatic birds and may mutate to high pathogenicity when they cross into poultry and cause debilitating Newcastle disease. This study characterised AMPV-1 isolated from fresh faecal droppings from wild aquatic birds roosting sites in Uganda. Fresh faecal samples from wild aquatic birds at several waterbodies in Uganda were collected and inoculated into 9–10-day-old embryonated chicken eggs. After isolation, the viruses were confirmed as APMV-1 by APMV-1-specific polymerase chain reaction (PCR). The cleavage site of the fusion protein gene for 24 representative isolates was sequenced and phylogenetically analysed and compared with representative isolates of the different APMV-1 genotypes in the GenBank database. In total, 711 samples were collected from different regions in the country from which 72 isolates were recovered, giving a prevalence of 10.1%. Sequence analysis of 24 isolates revealed that the isolates were all lentogenic, with the typical 111GGRQGR’L117 avirulent motif. Twenty-two isolates had similar amino acid sequences at the cleavage site, which were different from the LaSota vaccine strain by a silent nucleotide substitution T357C. Two isolates, NDV/waterfowl/Uganda/MU150/2011 and NDV/waterfowl/Uganda/MU186/2011, were different from the rest of the isolates in a single amino acid, with aspartate and alanine at positions 124 and 129, respectively. The results of this study revealed that Ugandan aquatic birds indeed harbour APMV-1 that clustered with class II genotype II strains and had limited genetic diversity.

International audience | Avian paramyxovirus type-1 (APMV-1) viruses of the lentogenic pathotypes are often isolated from wild aquatic birds and may mutate to high pathogenicity when they cross into poultry and cause debilitating Newcastle disease. This study characterised AMPV-1 isolated from fresh faecal droppings from wild aquatic birds roosting sites in Uganda. Fresh faecal samples from wild aquatic birds at several waterbodies in Uganda were collected and inoculated into 9-10-day-old embryonated chicken eggs. After isolation, the viruses were confirmed as APMV-1 by APMV-1-specific polymerase chain reaction (PCR). The cleavage site of the fusion protein gene for 24 representative isolates was sequenced and phylogenetically analysed and compared with representative isolates of the different APMV-1 genotypes in the GenBank database. In total, 711 samples were collected from different regions in the country from which 72 isolates were recovered, giving a prevalence of 10.1%. Sequence analysis of 24 isolates revealed that the isolates were all lentogenic, with the typical 111GGRQGR'L117 avirulent motif. Twenty-two isolates had similar amino acid sequences at the cleavage site, which were different from the LaSota vaccine strain by a silent nucleotide substitution T357C. Two isolates, NDV/waterfowl/Uganda/MU150/2011 and NDV/waterfowl/Uganda/MU186/2011, were different from the rest of the isolates in a single amino acid, with aspartate and alanine at positions 124 and 129, respectively. The results of this study revealed that Ugandan aquatic birds indeed harbour APMV-1 that clustered with class II genotype II strains and had limited genetic diversity.

Avian paramyxovirus type-1 (APMV-1) viruses of the lentogenic pathotypes are often isolated from wild aquatic birds and may mutate to high pathogenicity when they cross into poultry and cause debilitating Newcastle disease. This study characterised AMPV-1 isolated from fresh faecal droppings from wild aquatic birds roosting sites in Uganda. Fresh faecal samples from wild aquatic birds at several waterbodies in Uganda were collected and inoculated into 9–10-day-old embryonated chicken eggs. After isolation, the viruses were confirmed as APMV-1 by APMV-1-specific polymerase chain reaction (PCR). The cleavage site of the fusion protein gene for 24 representative isolates was sequenced and phylogenetically analysed and compared with representative isolates of the different APMV-1 genotypes in the GenBank database. In total, 711 samples were collected from different regions in the country from which 72 isolates were recovered, giving a prevalence of 10.1%. Sequence analysis of 24 isolates revealed that the isolates were all lentogenic, with the typical 111GGRQGR’L117 avirulent motif. Twenty-two isolates had similar amino acid sequences at the cleavage site, which were different from the LaSota vaccine strain by a silent nucleotide substitution T357C. Two isolates, NDV/waterfowl/Uganda/MU150/2011 and NDV/waterfowl/Uganda/MU186/2011, were different from the rest of the isolates in a single amino acid, with aspartate and alanine at positions 124 and 129, respectively. The results of this study revealed that Ugandan aquatic birds indeed harbour APMV-1 that clustered with class II genotype II strains and had limited genetic diversity.