Objective: The objective of the current study was to investigate the influence of probiotic Chrysonilia crassa in comparison with zinc bacitracin, commercial probiotic Bacillus subtilis, and herbal medicine waste on growth, intestinal microbiology, and blood indices of broilers.Materials and methods: Three hundreds of Lohmann day-old chicks were allocated to control diet (basal diet; CONT), basal diet with antibiotic zinc bacitracin (AZB), basal diet with B. subtilis (PROB), basal diet with C. crassa (PROC), and basal diet with herbal medicine waste (HERBW). Sample collections were conducted on day 34 of the experiment.Results: PROB showed greater (P<0.05) body weight than CONT chicks. Leukocytes and lymphocytes numbers were higher (P<0.05) in HERBW than in birds of other treatments. Compared to CONT and HERBW, PROC birds showed higher (P<0.05) level of vaccine titer to Newcastle disease virus. CONT had lower (P<0.05) and higher (P<0.05) total protein and globulin, and the ratio of albumin to globulin (A/G ratio) in serum, respectively, compared to other chicks. Higher level (P<0.05) of albumin was observed in PROB relative to CONT and PROC. Lower uric acid (P<0.05) was seen in PROC when compared with CONT and PROB. PROC had higher (P<0.05) aspartate aminotransferase than AGP, PROB, and HERBW. Ileal coliform was decreased (P<0.05) in PROB and PROC, relative to CONT and HERBW.Conclusion: Probiotics were capable of improving the growth, immune responses, and intestinal bacterial populations of broilers. The effects of probiotics C. crassa corresponded to that of commercial probiotic B. subtilis and antibiotic growth promoters. http://doi.org/10.5455/javar.2018.e284

Objective: The objective of the current study was to investigate the influence of probiotic Chrysonilia crassa in comparison with zinc bacitracin, commercial probiotic Bacillus subtilis, and herbal medicine waste on growth, intestinal microbiology, and blood indices of broilers. Materials and methods: Three hundreds of Lohmann day-old chicks were allocated to control diet (basal diet; CONT), basal diet with antibiotic zinc bacitracin (AZB), basal diet with B. subtilis (PROB), basal diet with C. crassa (PROC), and basal diet with herbal medicine waste (HERBW). Sample collections were conducted on day 34 of the experiment. Results: PROB showed greater (P [J Adv Vet Anim Res 2018; 5(3.000): 332-342]

Objective: The present study was initiated to ascertain the level of shedding of salmonellae by dogs in Makurdi area and to highlight the risk of infection for dog-owners. Materials and Methods: Rectal swabs from 200 dogs from different locations in the study area were examined in the study. The samples were cultured for salmonellae using RappaportVassiliadis enrichment broth (Oxoid) and brilliant green agar (Oxoid). Suspected Salmonella isolates were serologically identified. Results: Overall, Salmonellae organisms were isolated from 11 (5.5%) of the 200 dogs sampled. Prevalence rates of 5.6% and 4.5% were recorded for apparently healthy and clinically sick dogs, respectively. Salmonella was respectively isolated from 4.1% to 9.1% of male and female dogs. Dogs aged 4 years and above recorded the highest prevalence rate. The study revealed a low prevalence rate in Nigerian local breed (mongrels) and high prevalence rates in exotic breeds of dogs. Conclusion: The isolation of salmonellae in apparently healthy and clinically sick dogs in this study indicates a carrier status which may constitute a serious problem in disease control in the study area. The lower prevalence rate of Salmonella infection in mongrels could be an indication of resistance to Salmonella in local breeds of dogs and should generate interest in research in the pathogenicity and pathogenesis of salmonellae in mongrels. http://doi.org/10.5455/javar.2018.e291

Objective: The present study was initiated to ascertain the level of shedding of salmonellae by dogs in Makurdi area and to highlight the risk of infection for dog-owners. Materials and Methods: Rectal swabs from 200 dogs from different locations in the studyarea were examined in the study. The samples were cultured for salmonellae using Rappaport-Vassiliadis enrichment broth (Oxoid) and brilliant green agar (Oxoid). Suspected Salmonella isolates were serologically identified. Results: Overall, Salmonellae organisms were isolated from 11 (5.5%) of the 200 dogs sampled. Prevalence rates of 5.6% and 4.5% were recorded for apparently healthy and clinically sick dogs, respectively. Salmonella was respectively isolated from 4.1% to 9.1% of male and female dogs. Dogs aged 4 years and above recorded the highest prevalence rate. The study revealed a low prevalence rate in Nigerian local breed (mongrels) and high prevalence rates in exotic breeds of dogs. Conclusion: The isolation of salmonellae in apparently healthy and clinically sick dogs in this study indicates a carrier status which may constitute a serious problem in disease control in the study area. The lower prevalence rate of Salmonella infection in mongrels could be an indication of resistance to Salmonella in local breeds of dogs and should generate interest in research in the pathogenicity and pathogenesis of salmonellae in mongrels. [J Adv Vet Anim Res 2018; 5(4.000): 405-409]

Objective: The objective of this study was to determine the prevalence, geographical distribution, and main risk factors for peste des petits ruminants (PPR) in the Republic of Chad. Materials and methods: A total of 3,546 sera collected from unvaccinated small ruminants including 1,699 goats and 1,847 sheep in 19 of the 23 regions in Chad were randomly sampled. The competitive enzyme-linked immunosorbent assay technics were used for serological analysis. Results: The overall seroprevalence at the individual level was 52.9%±1.6% (48.9% for goats and 56.2% for sheep). Seroprevalence observed in the Chari Baguirmi, Ouaddaï, and N’Djamena regions was significantly higher than those in the other regions. Transhumant herds are the most exposed than the sedentary ones. Older animals were more affected than the young ones. Kababich sheep are the most affected than other breeds. Conclusion: This study has shown that the PPR virus is circulating in the Republic of Chad. In view of the results obtained, the disease is enzootic in the country. Epidemiological information obtained including seroprevalence rate, risk factors (sex, breed, age, and mode of rearing), and geographical distribution will help to define an appropriate strategy for PPR control in the Republic of Chad. http://doi.org/10.5455/javar.2018.e293

Objective: The objective of this study was to determine the prevalence, geographical distribution, and main risk factors for peste des petits ruminants (PPR) in the Republic of Chad. Materials and methods: A total of 3,546 sera collected from unvaccinated small ruminants including 1,699 goats and 1,847 sheep in 19 of the 23 regions in Chad were randomly sampled. The competitive enzyme-linked immunosorbent assay technics were used for serological analysis. Results: The overall seroprevalence at the individual level was 52.9%±1.6% (48.9% for goats and 56.2% for sheep). Seroprevalence observed in the Chari Baguirmi, Ouaddaï, and NDjamena regions was significantly higher than those in the other regions. Transhumant herds are the most exposed than the sedentary ones. Older animals were more affected than the young ones. Kababich sheep are the most affected than other breeds. Conclusion: This study has shown that the PPR virus is circulating in the Republic of Chad. In view of the results obtained, the disease is enzootic in the country. Epidemiological information obtained including seroprevalence rate, risk factors (sex, breed, age, and mode of rearing), and geographical distribution will help to define an appropriate strategy for PPR control in the Republic of Chad. [J Adv Vet Anim Res 2018; 5(4.000): 420-425]

Objective: The aim of this research is to study the effectiveness of mosquito magnet (MM) for reducing mosquitoes (Diptera) populations in coastal areas. Materials and methods: The study sites are in the coastal area of Samut Songkhram province, Thailand, which is divided into two locations; one that is 2 km and another that is 4 km in distance from the sea. We used the Mosquito Magnet® Independence (MMI) trap for effective field testing in Samut Songkhram Province, Thailand. Traps were placed 100 m away from the house (one trap per location) and mosquitoes were collected at night from 6 PM to 6 AM during September and October 2017 (30 days). Results: A total of 2,561 adult mosquitoes, including Anopheles epiroticus Linton & Harbach, Culex quinquefasciatus Say, Cx. sitiens Wiedmann, and Cx. gelidus Theobald were collected by MMI. At a 2-km distance from the sea were captured more mosquitoes per night more than at a 4-km distance (63.63 ± 42.30 vs. 21.70 ± 12.42). The comparison of effectiveness of MMI in two locations of the coastal area was shown to have a statistically significant difference (p < 0.05) and analysis of the correlation between the number of mosquitoes caught in coastal areas, including at a 2- and 4-km distance from the sea, accounting for weather factors, we found that the effectiveness of MMI was not correlated with weather (p > 0.05). Conclusion: Overall, this study demonstrated that MM can be used to control mosquitoes in coastal areas with high efficiency, especially 2 km away from the sea. http://doi.org/10.5455/javar.2018.e294

Objective: The aim of this research is to study the effectiveness of mosquito magnet (MM) for reducing mosquitoes (Diptera) populations in coastal areas. Materials and methods: The study sites are in the coastal area of Samut Songkhram province, Thailand, which is divided into two locations; one that is 2 km and another that is 4 km in distance from the sea. We used the Mosquito Magnet® Independence (MMI) trap for effective field testing in Samut Songkhram Province, Thailand. Traps were placed 100 m away from the house (one trap per location) and mosquitoes were collected at night from 6 PM to 6 AM during September and October 2017 (30 days). Results: A total of 2,561 adult mosquitoes, including Anopheles epiroticus Linton & Harbach, Culex quinquefasciatus Say, Cx. sitiens Wiedmann, and Cx. gelidus Theobald were collected by MMI. At a 2-km distance from the sea were captured more mosquitoes per night more than at a 4-km distance (63.63 ± 42.30 vs. 21.70 ± 12.42). The comparison of effectiveness of MMI in two locations of the coastal area was shown to have a statistically significant difference (p < 0.05) and analysis of the correlation between the number of mosquitoes caught in coastal areas, including at a 2- and 4-km distance from the sea, accounting for weather factors, we found that the effectiveness of MMI was not correlated with weather (p > 0.05). Conclusion: Overall, this study demonstrated that MM can be used to control mosquitoes in coastal areas with high efficiency, especially 2 km away from the sea. [J Adv Vet Anim Res 2018; 5(4.000): 426-431]

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]

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.