Molecular detection and genetic characterization of Mycoplasma gallisepticum and Mycoplasma synoviae in selected chicken breeds in South Africa.

BACKGROUND: The impact of chickens on maintaining the economy and livelihood of rural communities cannot be overemphasized. In recent years, mycoplasmosis has become one of the diseases that affect the success of South African chicken production. Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) are the most prevalent strains of Mycoplasma in South Africa. MG and MS are significant respiratory pathogens affecting the productivity of chickens. The present study aimed to molecularly detect using qPCR and characterize the presence of MG and MS using phylogenetic analysis. The phylogenetic analysis was utilized to clarify general evolutionary relationships between related taxa of different MG and MS observed in tracheal swabs from South African chicken breeds. METHODS: Forty-five tracheal swabs of the Lohmann Brown (n = 9), Rhode Island Red (n = 9), Ovambo (n = 9), Venda (n = 9), and Potchefstroom Koekoek (n = 9) breeds were collected from symptomatic chickens present in the commercial farm. To detect MG and MS, DNA was extracted from tracheal swabs and faecal samples, and qPCR was performed with a 16 s rRNA (310 bp) and vlhA (400 bp) gene fragment. Following the sequencing of all the amplicons, MG, and MS dendrograms showing the evolutionary relationships among the five South African chicken breeds and the GeneBank reference population were constructed. RESULTS: The qPCR revealed the presence of MG and MS in 22% (2/9) of the tracheal swab samples tested for MS only in Rhode Island Red breeds; 66.6% (6/9) and 33% (3/9) of the tested samples in Ovambo breeds; and 11.1% (1/9) and 44.4% (4/9) of the tested samples in Venda breeds. No MG or MS were detected in the Lohmann Brown or Potchefstroom Koekoek breed. Furthermore, qPCR revealed the presence of MG in pooled faecal samples from Lohmann Brown and Ovambo breeds. Eight different bacterial isolates were recognized from both samples. Four isolates were of the 16 s ribosomal ribonucleic acid (rRNA) gene (named PT/MG51/ck/00, PT/MG48/ck/00, PT/MG41/ck/00 and PT/MG71/ck/00) gene of Mycoplasma gallisepticum, and the other was Mycoplasma Synoviae variable lipoprotein hemagglutinin A (vlhA) gene (named PT/MSA22/ck/01, PT/MS41/ck/01, PT/MS74/ck/01 and PT/MS46/ck/01) which were available in GenBank. These isolates were successfully sequenced with 95-100% similarity to the isolates from the gene bank. CONCLUSION: The study revealed the presence of both MG and MS in the chicken breeds sampled. Furthermore, the different breeds of chicken were found to be susceptible to infection under the intensive or commercial management system. Therefore, continuous surveillance is encouraged to prevent the spread and outbreak of MG and MS in the poultry industry in South Africa.

Comparative genomics analysis and characterization of Shiga toxin-producing Escherichia coli O157:H7 strains reveal virulence genes, resistance genes, prophages and plasmids.

Escherichia coli O157:H7 is a foodborne pathogen that has been linked to global disease outbreaks. These diseases include hemorrhagic colitis and hemolytic uremic syndrome. It is vital to know the features that make this strain pathogenic to understand the development of disease outbreaks. In the current study, a comparative genomic analysis was carried out to determine the presence of structural and functional features of O157:H7 strains obtained from 115 National Center for Biotechnology Information database. These strains of interest were analysed in the following programs: BLAST Ring Image Generator, PlasmidFinder, ResFinder, VirulenceFinder, IslandViewer 4 and PHASTER. Five strains (ECP19-198, ECP19-798, F7508, F8952, H2495) demonstrated a great homology with Sakai because of a few regions missing. Five resistant genes were identified, however, Macrolide-associated resistance gene mdf(A) was commonly found in all genomes. Majority of the strains (97%) were positive for 15 of the virulent genes (espA, espB, espF, espJ, gad, chuA, eae, iss, nleA, nleB, nleC, ompT, tccP, terC and tir). The plasmid analysis demonstrated that the IncF group was the most prevalent in the strains analysed. The prophage and genomic island analysis showed a distribution of bacteriophages and genomic islands respectively. The results indicated that structural and functional features of the many O157:H7 strains differ and may be a result of obtaining mobile genetic elements via horizontal gene transfer. Understanding the evolution of O157:H7 strains pathogenicity in terms of their structural and functional features will enable the development of detection and control of transmission strategies.

Genetic parameter estimates for daily predicted gross feed efficiency and its association with energy-corrected milk in South African Holstein cattle.

Genetic parameters for daily predicted gross feed efficiency (pGFE) and energy corrected milk (ECM) in the first three parities of South African Holstein cattle were estimated by repeatability animal models. Data comprised of 11,068 test-day milk production records of 1,575 Holstein cows that calved between 2009 and 2019. Heritability estimates for pGFE were 0.12 ± 0.06, 0.09 ± 0.04 and 0.18 ± 0.05 in early, mid and late lactation, respectively. Estimates were moderate for primiparous (0.21 ± 0.05) and low for multiparous (0.10 ± 0.04) cows. Heritability and repeatability across all lactations were 0.14 ± 0.03 and 0.37 ± 0.03, respectively. Genetic correlations between pGFE in different stages of lactation ranged from 0.87 ± 0.24 (early and mid) to 0.97 ± 0.28 (early and late), while a strong genetic correlation (0.90 ± 0.03) was found between pGFE and ECM, across all lactations. The low to moderate heritability estimates for pGFE suggest potential for genetic improvement of the trait through selection, albeit with a modest accuracy of selection. The high genetic correlation of pGFE with ECM may, however, assist to improve accuracy of selection for feed efficiency by including both traits in multi-trait analyses. These genetic parameters may be used to estimate breeding values for pGFE, which will enable the trait to be incorporated in the breeding objective for South African Holstein cattle.

Prediction of dry matter intake and gross feed efficiency using milk production and live weight in first-parity Holstein cows.

Direct measurement of dry matter intake (DMI) presents a major challenge in estimating gross feed efficiency (GFE) in dairy cattle. This challenge can, however, be resolved through the prediction of DMI and GFE from easy-to-measure traits such as milk production (i.e. milk yield, energy-corrected milk (ECM), butterfat, protein, lactose) and live weight (LW). The main objective of this study was, therefore, to investigate the feasibility of predicting dry matter intake and gross feed efficiency for first-parity Holstein cows using milk production traits and LW. Data comprised of 30 daily measurements of DMI and milk production traits, and 25 daily LW records of a group of 100 first-parity Holstein cows, fed a total mixed ration. Gross feed efficiency was calculated as kg ECM divided by kg DMI. The initial step was to estimate correlations of milk production traits and LW with DMI and GFE, to identify the best potential predictors of DMI and GFE. Subsequently, a forward stepwise regression analysis was used to develop models to predict DMI and GFE from LW and milk production traits, followed by within-herd validations. Means for DMI, butterfat yield (BFY) and LW were 21.91 ± 2.77 kg/day, 0.95 ± 0.14 kg/day and 572 ± 15.58 kg/day, respectively. Mean GFE was 1.32 ± 0.22. Dry matter intake had positive correlations with milk yield (MY) (r = 0.32, p < 0.001) and LW (r = 0.76, p < 0.0001) and an antagonistic association with butterfat percent (BFP) (r = - 0.55, p < 0.001). On the other hand, GFE was positively associated with MY (r = 0.36, p < 0.001), BFP (r = 0.53, p < 0.001) and BFY (r = 0.83, p < 0.0001), and negatively correlated with LW (r = - 0.23, p > 0.05). Dry matter intake was predicted reliably by a model comprising of only LW and MY (R2 = 0.79; root mean squared error (RMSE) = 1.05 kg/day). A model that included BFY, MY and LW had the highest ability to predict GFE (R2 = 0.98; RMSE = 0.05). Live weight and BFY were the main predictor traits for DMI and GFE, respectively. The best models for predicting DMI and GFE were as follows: DMI (kg/day) = - 54.21 - 0.192 × MY (kg/day) + 0.146 × LW (kg/day) and GFE (kg/day) = 4.120 + 0.024 × MY (kg/day) + 1.000 × BFY (kg/day) - 0.008 × LW (kg/day). Thus, daily DMI (kg/day) and GFE can be reliably predicted from LW and milk production traits using these developed models in first-parity Holstein cows. This presents a big promise to generate large quantities of data of individual cow DMI and GFE, which can be used to implement genetic improvement of feed efficiency.

Novel single nucleotide polymorphisms in the heat shock protein 70.1 gene in South African Nguni crossbred cattle.

High environmental temperatures are one of the main causes of reduced productivity and reproduction in livestock. In an endeavour to counteract the effects of high temperature, a special class of proteins known as heat shock proteins function to alleviate heat stress in the cells. In this study, two regions (3'- and 5'-UTR) of the heat shock protein 70.1 (HSP70.1) gene were studied in Nguni crossbred cattle. Subsequently, the population genetic structure was elucidated. The 5'-UTR contained the most polymorphisms with 46 and 67 SNPs, while the 3'-UTR contained 7 and 16 SNPs in the Umzimkulu and Port Shepstone populations, respectively. The T64G polymorphism had the greatest frequency of all returned SNPs in the 3'-UTR; it was fixed for the Umzimkulu population (allele frequency = 1.00) and was nearing fixation in the Port Shepstone population (allele frequency = 0.979). In the 5'-UTR, the cytosine insertion at position 1110 was fixed for both populations. These polymorphisms are presumed to play a major role in the high thermotolerance exhibited in Nguni crossbred cattle. Partitioning of genetic variation displayed that the majority of the variation (96%) was within populations, whereas only 4% of the variation was due to population genetic differentiation. A total of 22 haplotypes defined the 5'-UTR while the 3'-UTR contained 4. In conclusion, this study demonstrated that Nguni crossbred cattle in the KwaZulu-Natal province of South Africa are clustered into two genetic groups based on the HSP70.1 gene. The findings of this research will provide future directions on the identification of important SNPs within the HSP70.1 gene in South African indigenous cattle because this is one of the first of such studies.

Population genetic structure and maternal lineage of South African crossbred Nguni cattle using the cytochrome b gene in mtDNA.

The Nguni cattle breed predominates South Africa and is endowed with traits favourable against environmental stressors such as heat stress and resistance to diseases. Interventions to improve production have led to the erosion of the genetic integrity of local breeds and the introduction of exotic breeds has proved ineffective as they fail to perform well due to different climatic conditions and production systems. In this study, the genetic structure and genetic lineage of Nguni crossbreds from 6 populations were assessed using the mitochondrial cytochrome b gene. Twelve polymorphic sites were detected resulting in 11 haplotypes with haplotype and nucleotide diversities of 0.550 ± 0.135 and 0.0019 ± 0.0011, respectively. Only 2 of the 6 populations displayed recent population expansion events, whereas the majority adhered to neutral evolution. The basal haplotype contained approximately 60% of the studied populations and there were four unique haplotypes that were revealed. A possible Nguni descript haplotype was uncovered, and this haplotype was found in all populations but was however devoid of individuals from around the world. The genetic structure of the populations was rather low (average pairwise FST = 0.066 and Slatkins FST = 0.094), and approximately 96% of the total genetic variation was accounted for by differences within populations. Phylogenetic analyses supported the clustering of all the samples within the Bos taurus clade and no Bos indicus haplotype was detected. Furthermore, no intermediate haplotype of taurine and indicine was detected. Overall, the maternal lineage of the crossbreds points to a taurine origin and the low genetic diversity depicts the retention of the Nguni genetic pool and possibly its superior adaptive traits.

Molecular detection of virulence genes in Staphylococcus aureus isolated from wild pigeons (Columba domestica livia) in KwaZulu-Natal in South Africa.

The current study aimed to determine virulence determinants among S. aureus isolated from wild pigeons and houseflies around hospital areas in the Greater Durban area, South Africa. Following enrichment and bacterial growth, DNA extraction using the boiling method was performed. Overall, 57 out of 252 samples (22.6%) were positive for S. aureus. Six known virulence genes were tested, where five known virulence determinants were positive and none of the S. aureus isolates were positive to coagulase (coa) gene. The highest prevalence rates were found in the genes encoding haemolysins, with the hla and hld genes having 8 (14%) and 9 (15.8%) positive isolates respectively. The sea, LukS/F-PV, and spa genes had 5 (8.8%), 4 (7%), and 2 (3.5%) positive isolates respectively. These results demonstrated the detection of pathogenic S. aureus from hospital environment in Durban, South Africa which may account for the emergence staphylococcal infections. The findings of the present study highlights the significant role of wild pigeons and houseflies as potenital infectious disease vectors in a One Health context.

Molecular detection of multidrug and methicillin resistance in Staphylococcus aureus isolated from wild pigeons (Columba livia) in South Africa.

Staphylococcus aureus is an important human and veterinary pathogen. The present study aimed to determine the prevalence of antibiotic resistance among S. aureus isolated from samples obtained from free-flying wild pigeons and houseflies from different locations surrounding a local hospital in the Greater Durban area in KwaZulu-Natal Province, South Africa. Environmental fecal samples were obtained from wild pigeons that inhabits the grounds of a local public hospital located on the South Beach area, Durban, South Africa. Housefly samples were collected from three different locations (Kenneth Stainbank Nature Reserve, Montclair/Clairwood, and Glenwood/Berea) in the greater Durban area, all within a close proximity to the hospital. Following enrichment, identification, and antimicrobial resistance profiling, S. aureus isolates were subjected to DNA extraction using the boiling method. It was found that 57 out of 252 samples (22.62%) were positive for S. aureus. The Kirby-Bauer disk diffusion method of antibiotic susceptibility testing was performed and revealed that antibiotic resistance rates to penicillin and rifampicin were the most common, with both returning 48 (84.2%) out of the 57 S. aureus isolates being resistant to penicillin and rifampicin. Antibiotic resistance rates to clindamycin, linezolid, erythromycin, tetracycline, cefoxitin, and ciprofloxacin were 82.5%, 78.9%, 73.7%, 63.2%, 33.3%, and 15.8% respectively. Antibiotic resistance genes were detected using primer-specific PCR and it was found that the prevalence rates of tetM, aac(6')-aph(2″), mecA, tetK, ermc, and blaZ genes were 66.7%, 40.4%, 40.4%, 38.6%, 24.6%, and 3.51% respectively. Statistical analysis revealed significant (p < 0.05) relationships between the tetM, aac(6')-aph(2″), and ermC genes and all parameters tested. A significant correlation between the aac(6')-aph(2″) gene and the tetM (0.506) and ermC (-0.386) genes was identified. It was found that 23 (40.3%) S. aureus isolates were mecA positive, of which 10 (52.6%) out of 19 cefoxitin-resistant isolates were mecA positive and 13 (35.1%) out of 37 cefoxitin-sensitive isolates were mecA positive. The results of the present study demonstrated the detection of methicillin and multidrug resistant S. aureus isolated from samples obtained from wild pigeons and houseflies in the surroundings of a local public hospital in the Greater Durban area in South Africa. The findings of the study may account for the emergence of multidrug-resistant staphylococcal infections. The findings highlight the significant role of wild pigeons and houseflies in the spread of drug-resistant pathogenic S. aureus including MRSA. The conclusions of the present study highlight the improtant role of wildlife and the environment as interconnected contributors of One Health.

Draft genome sequences of Bacillus licheniformis strains MEZBL63 and MEZBL64 harboring the lichenysin toxin operon isolated from livestock in South Africa.

Here, we report the draft genome sequences of two Bacillus licheniformis strains harboring the lichenysin operon that were isolated from healthy goat and horse in South Africa. The genomes were sequenced using Illumina MiSeq and had a length of 4,152,826 and 4,110,075 bp, respectively, with a G + C content of 46%.

Genetic trends for objectively measured and subjectively assessed traits in a Dorper sheep flock.

Genetic trends were constructed to monitor the genetic change for subjectively assessed and objectively measured traits using data emanating from complete records from the National Small Stock Improvement Scheme database and performance records accumulated by a single breeder over a period of 24 years. The objectively measured production traits considered were weaning weight, post-weaning weight (PWW), yearling weight, average daily weight gain to weaning (ADGW), average daily weight gain during post-weaning phase (ADGPW) and average daily weight gain up to yearling age. The subjectively assessed traits (scored on a five-point scale) were conformation, fat distribution, size, type and colour. Direct genetic trends for live weight and growth traits (with the exception of ADGPW) were positive. All the objectively measured traits where maternal effects were significant, except PWW, registered small declines in maternal breeding values. The fastest genetic progress was attained by ADGW, which amounted to 0.29 % of the overall phenotypic mean per annum. Conformation and type exhibited positive but slow increments in direct breeding values at an equivalent annual rate amounting to 0.12 and 0.09 % of the overall phenotypic mean, respectively. Size demonstrated a negative genetic trend of -0.14 % of the overall phenotypic mean per annum. Genetic trends for fat distribution and colour were negligible. It was concluded that breeders should focus more on the performance recording of objective traits as they are likely to respond favourably to selection pressure.

Antimicrobial resistance and whole genome sequencing of novel sequence types of Enterococcus faecalis, Enterococcus faecium, and Enterococcus durans isolated from livestock.

The emergence of antimicrobial-resistant, livestock-associated Enterococcus faecalis represents a public health concern. Here, we report the isolation, molecular detection of virulence and antimicrobial resistance determinants, in addition to the phylogenetic analyses of 20 Enterococcus species using whole genome sequencing analysis of 15 Enterococcus faecalis strains including six strains of three novel sequence types, three Enterococcus faecium and two Enterococcus durans. All strains were isolated from food chain animals in South Africa. Enterococcus strains were isolated on bile aesculin azide agar, followed by identification using MALDI-TOF MS analysis. Antibiotic susceptibility testing was performed using the Kirby-Bauer disk diffusion method. The genomic DNA of the isolates was extracted and sequencing was performed using the Illumina MiSeq platform. Sequence reads were trimmed and de novo assembled. The assembled contigs were analyzed for antimicrobial resistance genes and chromosomal mutations, extra-chromosomal plasmids, and multi-locus sequence type (MLST). Multidrug antimicrobial resistance genes conferring resistance to aminoglycosides (ant(6)-Ia, aph(3')-IIIa, sat4, and spw), lincosamides (lnu(B), lsa(A), and lsa(E)), macrolides (erm(B)), trimethoprim (dfrG) and tetracyclines (tet(L) and tet(M)) were identified. Plasmid replicons were detected in seven E. faecalis and three E. faecium isolates. The sequence type (ST) of each isolate was determined using the Enterococcus PubMLST database. Ten STs were identified in the collection, three of which (ST1240, ST1241, and ST1242) have not been previously reported and are described in the present study for the first time. To compare the sequenced strains to other previously sequenced E. faecalis strains, assembled sequences of E. faecalis from livestock were downloaded from the PubMLST database. Core genome-based phylogenetic analysis was performed using ParSNP. The detection of multiple drug-resistance in Enterococcus including E. faecalis and E. faecium highlights the significance of genomic surveillance to monitor the spread of antimicrobial resistance in food chain animals. In addition, the genome sequences of Enterococcus strains reported in the present study will serve as a reference point for future molecular epidemiological studies of livestock-associated and antibiotic-resistant E. faecalis in Africa. In addition, this study enables the in-depth analysis of E. faecalis genomic structure, as well as provides valuable information on the phenotypic and genotypic antimicrobial resistance, and the pathogenesis of livestock-associated E. faecalis and E. faecium.

Draft Genome Sequence of Listeria innocua Strain MEZLIS29, Isolated from a Cow in South Africa.

Here, we report the draft genome sequence of Listeria innocua strain MEZLIS29, which was isolated from a healthy cow in Flagstaff, Eastern Cape Province, South Africa. The genome was sequenced using the Illumina MiSeq platform and had a length of 2,805,865 bp, with a G+C content of 37.5% and 2,783 coding DNA sequences, 58 tRNAs, 4 noncoding RNAs, and 8 rRNA genes.

Review of Escherichia coli O157:H7 Prevalence, Pathogenicity, Heavy Metal and Antimicrobial Resistance, African Perspective.

Escherichia coli O157:H7 is an important food-borne and water-borne pathogen that causes hemorrhagic colitis and the hemolytic-uremic syndrome in humans and may cause serious morbidity and large outbreaks worldwide. People with bloody diarrhea have an increased risk of developing serious complications such as acute renal failure and neurological damage. The hemolytic-uremic syndrome (HUS) is a serious condition, and up to 50% of HUS patients can develop long-term renal dysfunction or blood pressure-related complications. Children aged two to six years have an increased risk of developing HUS. Clinical enteropathogenic Escherichia coli (EPEC) infections show fever, vomiting, and diarrhea. The EPEC reservoir is unknown but is suggested to be an asymptomatic or symptomatic child or an asymptomatic adult carrier. Spreading is often through the fecal-oral route. The prevalence of EPEC in infants is low, and EPEC is highly contagious in children. EPEC disease in children tends to be clinically more severe than other diarrheal infections. Some children experience persistent diarrhea that lasts for more than 14 days. Enterotoxigenic Escherichia coli (ETEC) strains are a compelling cause of the problem of diarrheal disease. ETEC strains are a global concern as the bacteria are the leading cause of acute watery diarrhea in children and the leading cause of traveler's diarrhea. It is contagious to children and can cause chronic diarrhea that can affect the development and well-being of children. Infections with diarrheagenic E. coli are more common in African countries. Antimicrobial agents should be avoided in the acute phase of the disease since studies showed that antimicrobial agents may increase the risk of HUS in children. The South African National Veterinary Surveillance and Monitoring Programme for Resistance to Antimicrobial Drugs has reported increased antimicrobial resistance in E. coli. Pathogenic bacterial strains have developed resistance to a variety of antimicrobial agents due to antimicrobial misuse. The induced heavy metal tolerance may also enhance antimicrobial resistance. The prevalence of antimicrobial resistance depends on the type of the antimicrobial agent, bacterial strain, dose, time, and mode of administration. Developing countries are severely affected by increased resistance to antimicrobial agents due to poverty, lack of proper hygiene, and clean water, which can lead to bacterial infections with limited treatment options due to resistance.

Molecular Surveillance and Dissemination of Klebsiella pneumoniae on Frequently Encountered Surfaces in South African Public Hospitals.

Bacteria that cause life-threatening illnesses in humans are also capable of contaminating hospital surfaces, thus pose as a potential source of infection. This study aimed to investigate the prevalence, genetic diversity, virulence, and antibiotic resistance profile of Klebsiella pneumoniae in South Africa. In a nonoutbreak setting involving four public hospitals, 777 samples were collected in three different wards from 11 different sites. Phenotypic and genotypic methods were used for isolation and identification. The Kirby-Bauer disk-diffusion method was used to examine antibiotic resistance followed by the combination disk method to characterize extended-spectrum ß-lactamases (ESBLs). Antibiotic resistance and virulence genes were screened using PCR and clonality was investigated using enterobacterial repetitive intergenic consensus (ERIC)-PCR. Seventy-five (10%) K. pneumoniae isolates were recovered. These isolates were obtained from all four hospitals and all three wards involved. However, only six frequently touched surfaces were contaminated. Thirty (40%) isolates were characterized as ESBLs showing high resistance to antibiotics and mostly harboring the blaCTX-M group one gene. Virulence genes were highly prevalent among all the isolates. ERIC-PCR showed that the isolates recovered from different sites within the same hospital were genetically similar. The study highlighted that K. pneumoniae can contaminate various surfaces and this persistence allows for the dissemination of bacteria within the hospital environment. The information from this study can assist hospitals to evaluate and improve current infection prevention and control interventions in place.

Ticks of domestic animals in Lesotho: Morphological and molecular characterization.

A total of 3311 tick specimens were randomly collected from domestic animals including cattle, sheep, goats, horses, donkeys, and dogs from Lesotho districts namely, Berea, Butha-Buthe, Leribe, Mafeteng, Maseru, Mohale's Hoek, Mokhotlong, Qacha's Nek, Quthing and Thaba Tseka. Tick species were identified morphologically and verified by amplification and sequencing of the CO1 and 18S rRNA genes. Nine species were identified under different genera namely, Haemaphysalis elliptica 0.1% (n = 2), Hyalomma rufipes 2.6% (n = 87), Hy. truncatum 1.2% (n = 41), Otobius megnini 13.6% (n = 451), Rhipicephalus appendiculatus 0.1% (n = 3), Rhipicephalus decoloratus 9.3% (n = 308), Rhipicephalus evertsi evertsi 65.1% (n = 2156), Rhipicephalus glabroscutatum 1.3% (n = 43) and Rhipicephalus microplus 6.6% (n = 220). There was a significant difference at p = 6.2E-06 (ꭓ2 = 1.923, df = 7) in the distribution of tick species and their abundance p = 0.04 (ꭓ2 = 1.923, df = 7) from each population. The CO1 and 18S rRNA sequences matched the morphological determinations on the NCBI database and clustered with relevant species on the phylogenetic tree. Genetic analysis of CO1 and 18S rRNA provided very strong support for monophyly of the Rhipicephalinae and Ornithodorinae complexes. Both CO1 and 18S rRNA are useful genetic markers for the specific and generic characterization of tick species in Lesotho and elsewhere. This is the first scientific publication of tick species occurring in Lesotho.

Genetic Variation and Population Differentiation in the Bovine Lymphocyte Antigen DRB3.2 Locus of South African Nguni Crossbred Cattle.

The bovine lymphocyte antigen (BoLA-DRB3) gene is an important region that codes for glycoproteins responsible for the initiation of an immune response. BoLA-DRB3 alleles have been demonstrated to be associated with disease resistance/tolerance. Therefore, great genetic diversity is correlated with better adaptation, fitness, and robustness. The current study was conducted to assess the population genetic structure of the BoLA-DRB3 gene in Nguni crossbred cattle using polymerase chain reaction-sequence based typing (PCR-SBT). High genetic diversity was detected, with 30 alleles, 11 of which are novel to the study. Alleles DRB3*0201, DRB3*0701, DRB*0901, and DRB*1601 were present in all populations and accounted for nearly around 50% of all observed alleles. A mean genetic diversity (HE) of 0.93 was detected. The high overall genetic diversity is possibly associated with pathogen-assisted selection and heterozygote advantage. Such high diversity might explain the hardiness of the Nguni crossbred cattle to the Southern African region. Low population genetic structure was identified (FST = 0.01), suggesting possible gene flow between populations and retention of similar alleles. The study was undertaken to bridge the dearth of such studies in South African breeds and it is imperative for effective sustainability of indigenous breeds and the implementation of effective breeding strategies.

A descriptive study of parasites detected in ticks of domestic animals in Lesotho.

Ticks are medically important and significant vectors of diseases affecting livestock, humans, and companion animals than any other arthropod vectors. In the absence of information on the relationship of tick species and piroplasms parasites in Lesotho, the current study was aimed at detecting piroplasms parasites of economic importance from ticks of domestic animals. A total of 322 pooled tick DNA samples were subjected to PCR screening for the presence of piroplasms. The overall infection rate of piroplasms was 7% with Babesia bigemina at 3.4% (11/322), B. bovis 0.3% (1/322), B. ovis 2.8% (9/322) and 0.6% (2/322) for B. motasi. DNA extracted from the Lesotho Rhipicephalus decoloratus and R. evertsi evertsi tested positive for the presence of B. bigemina with a 15% and 3% infection rate, respectively. Otobius megnini tested positive for only B. bovis at a 12.5% infection rate. Rhipicephalus e. evertsi was the only tick species PCR positive for ovine babesiosis with 3.2% for B. ovis and 0.7% for B. motasi. Equine piroplasm (Theileria equi and B. caballi) and Theileria (T. parva and T. ovis) parasites were not detected in the current study. The PCR-positive samples were confirmed by direct sequencing of the product. This study is the first to report on a relationship of Babesia parasites with tick species in Lesotho and it is evident that vector-borne diseases are present in ticks of domestic animals in this country. Research findings in this study require a joint effort from both veterinary and medical sectors to unite and conduct more epidemiological studies of tick-borne diseases in both animals and humans and to also determine the role played by tick species in the transmission of the detected parasites in domestic animals of Lesotho. This information provides a baseline knowledge of important piroplasms parasites and raising awareness of their prevalence in Lesotho.

Genomic Characterization of Antimicrobial Resistance in Food Chain and Livestock-Associated Salmonella Species.

The rising trend of antimicrobial resistance (AMR) by foodborne bacteria is a public health concern as these pathogens are easily transmitted to humans through the food chain. Non-typhoid Salmonella spp. is one of the leading foodborne pathogens which infect humans worldwide and is associated with food and livestock. Due to the lack of discovery of new antibiotics and the pressure exerted by antimicrobial resistance in the pharmaceutical industry, this review aimed to address the issue of antibiotic use in livestock which leads to AMR in Salmonella. Much attention was given to resistance to carbapenems and colistin which are the last-line antibiotics used in cases of multi drug resistant bacterial infections. In the present review, we highlighted data published on antimicrobial resistant Salmonella species and serovars associated with livestock and food chain animals. The importance of genomic characterization of carbapenem and colistin resistant Salmonella in determining the relationship between human clinical isolates and food animal isolates was also discussed in this review. Plasmids, transposons, and insertion sequence elements mediate dissemination of not only AMR genes but also genes for resistance to heavy metals and disinfectants, thus limiting the therapeutic options for treatment and control of Salmonella. Genes for resistance to colistin (mcr-1 to mcr-9) and carbapenem (blaVIM-1, blaDNM-1, and blaNDM-5) have been detected from poultry, pig, and human Salmonella isolates, indicating food animal-associated AMR which is a threat to human public health. Genotyping, plasmid characterization, and phylogenetic analysis is important in understanding the epidemiology of livestock-related Salmonella so that measures of preventing foodborne threats to humans can be improved.

Genotypic and Phenotypic Characterizations of Methicillin-Resistant Staphylococcus aureus (MRSA) on Frequently Touched Sites from Public Hospitals in South Africa.

The hospital environment acts as a reservoir in the transmission of pathogens, such as MRSA, which may cause hospital-acquired infections. This study aimed to ascertain the prevalence, genetic relatedness, antibiotic resistance, and virulence profile of MRSA on some frequently touched hospital sites in South Africa. A total of 777 swabs were randomly collected from 11 frequently touched sites in the hospital environment of three wards of four public hospitals in the KwaZulu-Natal province of South Africa. Isolation of S. aureus and confirmation were done using genotypic and phenotypic methods. Antibiotic susceptibility testing was performed using the Kirby-Bauer disk-diffusion method. MRSA isolates were determined by the presence of the mecA gene. Virulence and resistance genes were detected using a standard monoplex PCR assay. ERIC-PCR was conducted to evaluate the genetic relatedness. An overall prevalence of 12.7% for S. aureus isolates was obtained. Out of these, 89.9% (89/99) were confirmed to be MRSA. The sites with the highest prevalence were the occupied beds (16.2% (16/99)), unoccupied beds (16.2% (16/99)), patient files (14.1% (14/99)), ward phones (13.1% (13/99)), and nurses' tables (14.1% (14/99)). The virulence genes with the highest observed frequency were hld (87 (87.9%)) and LukS/F-PV (53 (53.5%)). The resistance genes with the highest frequency were the tetM and tetK genes detected in 60 (60.6%) and 57 (57.6%) isolates, respectively. The ERIC-PCR results obtained indicated a high level of genetic diversity; however, intraclonal (within a hospital) and interclonal (between hospitals) clusters of MRSA were observed. The study showed that MRSA can contaminate various surfaces, and this persistence allows for the dissemination of bacteria within the hospital environment. This highlights the need for improved infection prevention and control (IPC) strategies in public hospitals in the country to curb their potential transmission risks.

Occurrence, Antibiotic Resistance, Virulence Factors, and Genetic Diversity of Bacillus spp. from Public Hospital Environments in South Africa.

This study aimed to assess the molecular dissemination of Bacillus species in public hospitals in South Africa. The study conducted over 3 months during 2017 involved representative samples obtained from three wards (general ward, intensive care unit, and pediatric unit) from four public hospitals denoted as A (Central), B (Tertiary), C (Regional), and D (District). Swabs collected from 11 distinct hospital surfaces were screened using selective media, biochemical testing, and molecular methods. Overall, 17% (135/777) isolates were identified with a prevalence of 24% (32/135) for central, 33% (45/135) for tertiary, 27% (36/135) for regional, and 16% (22/135) for district hospital. Bacillus species were further confirmed to belong to Bacillus cereus (129/135; 96%) and Bacillus subtilis (6/135; 4%). Prevalence was similar across the wards, averaging 33.3% (45/135). The highest prevalence of Bacillus isolates was found on the drip stands (11.8%), sink (11.8%), ward phone (11.5%), and nurses' tables (10.3%). Minimum inhibitory concentration analyses revealed high resistance to ß-lactams, fluoroquinolones, and tetracyclines. The most common resistance genes detected were ermB (56%) and tetM (5%). Enterotoxin virulence genes hblA (77%) and hblD (88%) associated with the diarrheal syndrome were most detected; however, no ces genes (cereulide toxin) for emetic syndrome was found. The enterobacterial repetitive intergenic consensus PCR revealed considerable diversity at the different levels of health care, although the clonal spread of strains between the sites/wards within each specific hospital was revealed. The study highlighted the dissemination of drug-resistant Bacillus spp. in public hospital environments and calls for the design of optimal strategies to curb their spread.