To date, there is limited data about the genetic relationship of Escherichia coli between wild birds and cattle because these birds act as silent vectors for many zoonotic bacteria. This study aimed to elucidate the role of rooming wild birds in the vicinity of cattle farm in transmission of the same pathogenic E. coli variants, identifying their virulence, resistance traits and genetic similarities of fimH virulence gene. About 240 faecal/cloacal swabs were collected from both species and examined bacteriologically. Escherichia coli was yielded in 45.8% and 32.5%, respectively, of examined cattle and wild birds. The most prevalent detected E. coli serovar was O26. High tetracycline and chloramphenicol resistance were recorded; however, gentamycin and ciprofloxacin exhibited the highest sensitivity rates. Polymerase chain reaction (PCR) conserved genotypic resistance (tetA and blaCTX-M) and virulence attributes (fimH, stx1, eaeA and ompA) of E. coli isolates were discussed in detail. The fimH gene revealed 100% sequence similarity when comparing with different E. coli isolates globally and locally. Finally, a close genetic association of E. coli with both wild birds and cattle was detected, thus strengthening its role in the dissemination of the infection via environment. Prevention and conservative policy should be carried as E. coli constitute enormous significant zoonotic risks to livestock and animal workers. Also, further studies to the whole genome sequencing of fimH, other virulence and resistance genes of E. coli are recommended trying to limit the possibilities of co-infection and transfer among different species. Contribution: The current study recorded updated data about the critical infectious role of wild birds to livestock, including cattle farms in Egypt. It also delivered some recommendations for good hygienic practices in cattle farms which must be implemented for handling animal manure.

To date, there is limited data about the genetic relationship of Escherichia coli between wild birds and cattle because these birds act as silent vectors for many zoonotic bacteria. This study aimed to elucidate the role of rooming wild birds in the vicinity of cattle farm in transmission of the same pathogenic E. coli variants, identifying their virulence, resistance traits and genetic similarities of fimH virulence gene. About 240 faecal/cloacal swabs were collected from both species and examined bacteriologically. Escherichia coli was yielded in 45.8% and 32.5%, respectively, of examined cattle and wild birds. The most prevalent detected E. coli serovar was O26. High tetracycline and chloramphenicol resistance were recorded; however, gentamycin and ciprofloxacin exhibited the highest sensitivity rates. Polymerase chain reaction (PCR) conserved genotypic resistance (tetA and blaCTX-M) and virulence attributes (fimH, stx1, eaeA and ompA) of E. coli isolates were discussed in detail. The fimH gene revealed 100% sequence similarity when comparing with different E. coli isolates globally and locally. Finally, a close genetic association of E. coli with both wild birds and cattle was detected, thus strengthening its role in the dissemination of the infection via environment. Prevention and conservative policy should be carried as E. coli constitute enormous significant zoonotic risks to livestock and animal workers. Also, further studies to the whole genome sequencing of fimH, other virulence and resistance genes of E. coli are recommended trying to limit the possibilities of co-infection and transfer among different species. CONTRIBUTION: The current study recorded updated data about the critical infectious role of wild birds to livestock, including cattle farms in Egypt. It also delivered some recommendations for good hygienic practices in cattle farms which must be implemented for handling animal manure.

Neospora caninum is a coccidian parasite that occurs worldwide and is one of the most important causes of abortion, especially in cattle. However, no studies have been performed in Namibia to determine the N. caninum status in livestock. Therefore, this study aimed to determine the seroprevalence of N. caninum in cattle and the associated risk factors in the Khomas region of Namibia. A total of 736 sera were collected from cows in 32 farming establishments. These comprised 698 beef and 38 dairy cattle sera and were tested using a commercial enzyme-linked immunosorbent assay (ELISA) kit. Questionnaires were concurrently administered to determine possible risk factors associated with N. caninum seropositivity. A total of 42 sera were positive (all beef), giving an animal-level seroprevalence rate of 5.7%. Eight of the 32 establishments had at least one positive animal, giving a herd-level seroprevalence of 25%. There was no significant association between seropositivity and the presence of dogs, jackals, history of abortions, farm size, number of cattle or average annual rainfall. The establishments with moderate to high numbers of Feliformia were 9.8 times more likely to be seropositive to N. caninum than those with none to low levels of the former (p = 0.0245). The authors concluded that the seroprevalence level of N. caninum in the Khomas region was relatively low compared with other parts of the world and that the role of Feliformia in the epidemiology of bovine neosporosis needed to be further investigated. Contribution: Serological evidence of bovine neosporosis and the associated risk factors are reported in Namibia for the first time. This study contributes to the scientific body of knowledge on N. caninum in Africa, which is currently limited.

Neospora caninum is a coccidian parasite that occurs worldwide and is one of the most important causes of abortion, especially in cattle. However, no studies have been performed in Namibia to determine the N. caninum status in livestock. Therefore, this study aimed to determine the seroprevalence of N. caninum in cattle and the associated risk factors in the Khomas region of Namibia. A total of 736 sera were collected from cows in 32 farming establishments. These comprised 698 beef and 38 dairy cattle sera and were tested using a commercial enzyme-linked immunosorbent assay (ELISA) kit. Questionnaires were concurrently administered to determine possible risk factors associated with N. caninum seropositivity. A total of 42 sera were positive (all beef), giving an animal-level seroprevalence rate of 5.7%. Eight of the 32 establishments had at least one positive animal, giving a herd-level seroprevalence of 25%. There was no significant association between seropositivity and the presence of dogs, jackals, history of abortions, farm size, number of cattle or average annual rainfall. The establishments with moderate to high numbers of Feliformia were 9.8 times more likely to be seropositive to N. caninum than those with none to low levels of the former (p = 0.0245). The authors concluded that the seroprevalence level of N. caninum in the Khomas region was relatively low compared with other parts of the world and that the role of Feliformia in the epidemiology of bovine neosporosis needed to be further investigated. CONTRIBUTION: Serological evidence of bovine neosporosis and the associated risk factors are reported in Namibia for the first time. This study contributes to the scientific body of knowledge on N. caninum in Africa, which is currently limited.

Infection with Coxiella burnetii causes significant economic impact and poses zoonotic risk to people exposed to livestock, yet few studies in South Africa have assessed seroprevalence of C. burnetii infection and no information is available for goats. Very little information is available regarding risk factors and outcomes of C. burnetii infection in peri-urban farming areas where widespread mixing of ruminants occurs. This study estimated the seroprevalence of C. burnetii infection among communally farmed goats in an area adjacent to the densely populated Gauteng province. Sera were collected from 216 goats in 39 herds, and questionnaires were completed to establish management practices as potential risk factors. C. burnetii antibody testing was done by ELISA. Thirty two out of 216 goats tested positive for C. burnetii antibodies and the overall seroprevalence, adjusted for sampling weights and clustering, was 18.4% (95% confidence interval [CI]: 12.2% – 23.5%). The intraclass correlation coefficient was 0.06, indicating low-to-moderate clustering. Multiple logistic regression showed age was significantly associated with seropositivity, with higher seroprevalence in animals ≥ 19 months old (26%) than animals ≤ 6 months old (6%) (odds ratio [OR]: 6.6; p = 0.010). We concluded C. burnetii infection is common in goats in Moretele and a potential cause of abortion in goats and poses the potential zoonotic disease risk. Contribution: Despite the threats posed on animal health and productivity, scant information is published on C. burnetii in South Africa. This research established preliminary estimates of C. burnetii seroprevalence. The research is original from a South African perspective, relevant to Africa and focused on infectious disease in livestock.

Infection with Coxiella burnetii causes significant economic impact and poses zoonotic risk to people exposed to livestock, yet few studies in South Africa have assessed seroprevalence of C. burnetii infection and no information is available for goats. Very little information is available regarding risk factors and outcomes of C. burnetii infection in peri-urban farming areas where widespread mixing of ruminants occurs. This study estimated the seroprevalence of C. burnetii infection among communally farmed goats in an area adjacent to the densely populated Gauteng province. Sera were collected from 216 goats in 39 herds, and questionnaires were completed to establish management practices as potential risk factors. C. burnetii antibody testing was done by ELISA. Thirty two out of 216 goats tested positive for C. burnetii antibodies and the overall seroprevalence, adjusted for sampling weights and clustering, was 18.4% (95% confidence interval [CI]: 12.2% - 23.5%). The intraclass correlation coefficient was 0.06, indicating low-to-moderate clustering. Multiple logistic regression showed age was significantly associated with seropositivity, with higher seroprevalence in animals ≥ 19 months old (26%) than animals ≤ 6 months old (6%) (odds ratio [OR]: 6.6; p = 0.010). We concluded C. burnetii infection is common in goats in Moretele and a potential cause of abortion in goats and poses the potential zoonotic disease risk. CONTRIBUTION: Despite the threats posed on animal health and productivity, scant information is published on C. burnetii in South Africa. This research established preliminary estimates of C. burnetii seroprevalence. The research is original from a South African perspective, relevant to Africa and focused on infectious disease in livestock.

Cowdria polymorphic gene 1 (cpg1, Erum2510, ERUM_RS01380) has been shown to induce 30% and 100% protection in sheep immunised by deoxyribonucleic acid (DNA) prime combined with DNA boost and DNA prime combined with protein boost, respectively, against heartwater infection via needle challenge. To localise its antigenic regions for inclusion in a multi-epitope DNA vaccine against heartwater, Erum2510 was cleaved into five overlapping subfragments. These subfragments were expressed individually in an Escherichia coli host expression system and evaluated for their ability to induce proliferative responses, Th1 and Th2 cytokines (interferon gamma [IFN-γ] and interleukin 4 [IL-4]) via enzyme-linked immunospot (ELISpot), quantitative real time polymerase chain reaction (qRT-PCR) and flow cytometry. Recombinant (r)proteins 3 and 4 were shown to induce immunodominant Th1 and Th2 immune responses characterised by the secretion of effector cytokines IFN-γ and IL-4 in addition to differential messenger ribonucleic acid (mRNA) expression of tumour necrosis factor (TNF), IL-2, IL-1, IL-18, IL-10, transforming growth factor (TGF), granulocyte-macrophage colony-stimulating factor (GM-CSF) and inducible nitric oxide synthase (iNOS). Thirty-seven overlapping synthetic peptides (16 mer) spanning the lengths of these immunodominant rproteins were synthesised and assayed. A peptide pool comprising p9 and p10 derived from rprotein 3 induced a Th1-biased immune response. A peptide pool comprising p28 and p29 derived from rprotein 4 induced a mixed Th1 and Th2 immune response characterised by secretion of IFN-γ and differential mRNA expression of IL-1, IL-2, IL-10, IL-12, iNOS, TGF, TNF and GM-CSF. Only one of the peptides (p29) induced secretion of IL-4. Phenotypic analysis showed significant activation of cluster of differentiation 8+ (CD8+), cluster of differentiation 4+ (CD4+) and B+ lymphocyte populations. Findings suggest that Erum2510 rproteins and synthetic peptides can induce both cellular and humoral immune responses, thereby implicating their importance in protection against heartwater. Contribution: This study will facilitate the design of an effective multi-epitope DNA vaccine against heartwater that will contribute to control this economically important disease in sub-Saharan Africa and beyond.

Cowdria polymorphic gene 1 (cpg1, Erum2510, ERUM_RS01380) has been shown to induce 30% and 100% protection in sheep immunised by deoxyribonucleic acid (DNA) prime combined with DNA boost and DNA prime combined with protein boost, respectively, against heartwater infection via needle challenge. To localise its antigenic regions for inclusion in a multi-epitope DNA vaccine against heartwater, Erum2510 was cleaved into five overlapping subfragments. These subfragments were expressed individually in an Escherichia coli host expression system and evaluated for their ability to induce proliferative responses, Th1 and Th2 cytokines (interferon gamma [IFN-γ] and interleukin 4 [IL-4]) via enzyme-linked immunospot (ELISpot), quantitative real time polymerase chain reaction (qRT-PCR) and flow cytometry. Recombinant (r)proteins 3 and 4 were shown to induce immunodominant Th1 and Th2 immune responses characterised by the secretion of effector cytokines IFN-γ and IL-4 in addition to differential messenger ribonucleic acid (mRNA) expression of tumour necrosis factor (TNF), IL-2, IL-1, IL-18, IL-10, transforming growth factor (TGF), granulocyte-macrophage colony-stimulating factor (GM-CSF) and inducible nitric oxide synthase (iNOS). Thirty-seven overlapping synthetic peptides (16 mer) spanning the lengths of these immunodominant rproteins were synthesised and assayed. A peptide pool comprising p9 and p10 derived from rprotein 3 induced a Th1-biased immune response. A peptide pool comprising p28 and p29 derived from rprotein 4 induced a mixed Th1 and Th2 immune response characterised by secretion of IFN-γ and differential mRNA expression of IL-1, IL-2, IL-10, IL-12, iNOS, TGF, TNF and GM-CSF. Only one of the peptides (p29) induced secretion of IL-4. Phenotypic analysis showed significant activation of cluster of differentiation 8+ (CD8+), cluster of differentiation 4+ (CD4+) and B+ lymphocyte populations. Findings suggest that Erum2510 rproteins and synthetic peptides can induce both cellular and humoral immune responses, thereby implicating their importance in protection against heartwater. CONTRIBUTION: This study will facilitate the design of an effective multi-epitope DNA vaccine against heartwater that will contribute to control this economically important disease in sub-Saharan Africa and beyond.

Culicoides truuskae Labuschagne and Meiswinkel sp. n. is described and illustrated in both sexes from material collected in South Africa and Namibia. It is restricted to the xeric western margin of the subcontinent, occurring in Fynbos, Nama-Karoo and Succulent Karoo ecoregions in South Africa and Desert and Savanna ecoregions in Namibia experiencing 600 mm of rainfall annually. Culicoides truuskae sp. n. is part of the Afrotropical ‘plain-wing’ Culicoides in which the wing lacks a distinguishing pattern of light and dark spots; the diagnostic dark smudge that traverses wing cell r3 may result in C. truuskae sp. n. being misidentified as the sympatric but phyletically unrelated Culicoides herero (Enderlein) – (of the Similis group, subgenus Oecacta Poey). Additionally, this study is the first description of the male of C. herero. C. truuskae sp. n. and Culicoides coarctatus Clastrier and Wirth share similar characters in the male genitalia, although the two species are separable on wing pattern and female flagellum sensilla coeloconica (SCo) distribution. The breeding habitat and adult female blood-feeding preferences of C. truuskae sp. n. are not known. A maximum likelihood phylogenetic tree, using mitochondrial cytochrome c oxidase I (COI) sequence data, is provided to further clarify the relationship between C. truuskae sp. n., C. coarctatus and C. herero. Extensive light trap data, collected over 30 years, are used to map the distribution ranges of C. truuskae sp. n., C. coarctatus and C. herero in Southern Africa. Contribution: The description of this new species and the description of the male of C. herero increases our understanding of the diversity and distribution of Culicoides species in southern Africa.

Culicoides truuskae Labuschagne and Meiswinkel sp. n. is described and illustrated in both sexes from material collected in South Africa and Namibia. It is restricted to the xeric western margin of the subcontinent, occurring in Fynbos, Nama-Karoo and Succulent Karoo ecoregions in South Africa and Desert and Savanna ecoregions in Namibia experiencing < 600 mm of rainfall annually. Culicoides truuskae sp. n. is part of the Afrotropical 'plain-wing' Culicoides in which the wing lacks a distinguishing pattern of light and dark spots; the diagnostic dark smudge that traverses wing cell r3 may result in C. truuskae sp. n. being misidentified as the sympatric but phyletically unrelated Culicoides herero (Enderlein) - (of the Similis group, subgenus Oecacta Poey). Additionally, this study is the first description of the male of C. herero. C. truuskae sp. n. and Culicoides coarctatus Clastrier and Wirth share similar characters in the male genitalia, although the two species are separable on wing pattern and female flagellum sensilla coeloconica (SCo) distribution. The breeding habitat and adult female blood-feeding preferences of C. truuskae sp. n. are not known. A maximum likelihood phylogenetic tree, using mitochondrial cytochrome c oxidase I (COI) sequence data, is provided to further clarify the relationship between C. truuskae sp. n., C. coarctatus and C. herero. Extensive light trap data, collected over 30 years, are used to map the distribution ranges of C. truuskae sp. n., C. coarctatus and C. herero in Southern Africa. CONTRIBUTION: The description of this new species and the description of the male of C. herero increases our understanding of the diversity and distribution of Culicoides species in southern Africa.

Abortions in domestic ruminants cause significant economic losses to farmers. Determining the cause of an abortion is important for control efforts, but it can be challenging. All available diagnostic methods in the bacteriology laboratory should be employed in every case due to the many limiting factors (autolysis, lack of history, range of samples) that complicate the investigation process. The purpose of this study was to determine whether the recovery of diagnostically significant isolates from domestic ruminant abortion cases could be increased through the use of a combination of the existing aerobic culture and Brucella selective method with methods that are commonly recommended in the literature reporting abortion investigations. These methods are examination of wet preparations and impression smears stained by the modified Ziehl–Neelsen method, anaerobic, microaerophilic, Leptospira, Mycoplasma and fungal culture. Samples of placenta and aborted foetuses from 135 routine clinical abortion cases of cattle (n = 88), sheep (n = 25) and goats (n = 22) were analysed by the new combination of methods. In 46 cases, bacteria were identified as aetiological agents and in one case a fungus. Isolation of Brucella species increased to 7.4% over two years compared with the previous 10 years (7.3%), as well as Campylobacter jejuni (n = 2) and Rhizopus species (n = 1). Salmonella species (5.9%) and Trueperella pyogenes (4.4%) were also isolated more often. In conclusion, the approach was effective in removing test selection bias in the bacteriology laboratory. The importance of performing an in-depth study on the products of abortion by means of an extensive, combination of conventional culture methods was emphasised by increased isolation of Brucella abortus and isolation of C. jejuni. The combination of methods that yielded the most clinically relevant isolates was aerobic, microaerophilic, Brucella and fungal cultures.

Abortions in domestic ruminants cause significant economic losses to farmers. Determining the cause of an abortion is important for control efforts, but it can be challenging. All available diagnostic methods in the bacteriology laboratory should be employed in every case due to the many limiting factors (autolysis, lack of history, range of samples) that complicate the investigation process. The purpose of this study was to determine whether the recovery of diagnostically significant isolates from domestic ruminant abortion cases could be increased through the use of a combination of the existing aerobic culture and Brucella selective method with methods that are commonly recommended in the literature reporting abortion investigations. These methods are examination of wet preparations and impression smears stained by the modified Ziehl-Neelsen method, anaerobic, microaerophilic, Leptospira, Mycoplasma and fungal culture. Samples of placenta and aborted foetuses from 135 routine clinical abortion cases of cattle (n = 88), sheep (n = 25) and goats (n = 22) were analysed by the new combination of methods. In 46 cases, bacteria were identified as aetiological agents and in one case a fungus. Isolation of Brucella species increased to 7.4% over two years compared with the previous 10 years (7.3%), as well as Campylobacter jejuni (n = 2) and Rhizopus species (n = 1). Salmonella species (5.9%) and Trueperella pyogenes (4.4%) were also isolated more often. In conclusion, the approach was effective in removing test selection bias in the bacteriology laboratory. The importance of performing an in-depth study on the products of abortion by means of an extensive, combination of conventional culture methods was emphasised by increased isolation of Brucella abortus and isolation of C. jejuni. The combination of methods that yielded the most clinically relevant isolates was aerobic, microaerophilic, Brucella and fungal cultures.