Objective: This research aims to investigate the microbial diversity of Budu prepared from fresh and frozen fish from the Pariaman and Pasaman districts in West Sumatra Province, Indonesia, as well as provide basic information about Budu quality. Materials and methods: To obtain the bacterial microbial composition, deoxyribonucleic acid extraction was carried out using amplicon-sequencing of the 16S-rRNA gene in the V3–V4 region from two types of Budu and carried out in duplicate. Results: Budu prepared with fresh (Pariaman) or frozen (Pasaman) fish was dominated by Firmicutes (78.455%–92.37%) and Proteobacteria (6.477%–7.23%) phyla. The total microbial species in Budu from Pariaman were higher (227 species) than in Pasaman (153 species). The bacterial species found are Lentibacillus kimchi (1.878%–2.21%), Staphylococcus cohnii (0.597%– 0.70%), Peptostreptococcus russeli (0.00%–0.002%), Clostridium disporicum (0.073%–0.09%), Clostridium novyi (0.00%–0.01%), Nioella sediminis (0.00%–0.001%), and Shewanella baltica (0.00%–0.003%). Lentibacillus kimchi, S. cohnii, and C. disporicum are found in both Budu. Nioella sediminis and S. baltica are found in Budu Pariaman. Peptostreptococcus russeli and C. novyi were found in Budu Pasaman. Conclusion: Metagenomic analysis of Budu from different fish, Pariaman (fresh fish) and Pasaman (frozen fish) showed that the biodiversity of bacteria was barely different. Both Budu found lactic acid bacteria from the Enterococcaceae family, genus Vagococcus, and pathogenic bacteria, such as S. cohnii, P. russeli, C. disporicum, and S. baltica. The discovery of various species of pathogenic bacteria indicates that development is still needed in the Budu production process to improve Budu quality. J. Adv. Vet. Anim. Res., 10(4): 801–808, December 2023 http://doi.org/10.5455/javar.2023.j736

Objective: This research aims to investigate the microbial diversity of Budu prepared from fresh and frozen fish from the Pariaman and Pasaman districts in West Sumatra Province, Indonesia, as well as provide basic information about Budu quality. Materials and Methods: To obtain the bacterial microbial composition, deoxyribonucleic acid extraction was carried out using amplicon-sequencing of the 16S-rRNA gene in the V3–V4 region from two types of Budu and carried out in duplicate. Results: Budu prepared with fresh (Pariaman) or frozen (Pasaman) fish was dominated by Firmicutes (78.455%–92.37%) and Proteobacteria (6.477%–7.23%) phyla. The total microbial species in Budu from Pariaman were higher (227 species) than in Pasaman (153 species). The bacterial species found are Lentibacillus kimchi (1.878%–2.21%), Staphylococcus cohnii (0.597%–0.70%), Peptostreptococcus russeli (0.00%–0.002%), Clostridium disporicum (0.073%–0.09%), Clostridium novyi (0.00%–0.01%), Nioella sediminis (0.00%–0.001%), and Shewanella baltica (0.00%–0.003%). Lentibacillus kimchi, S. cohnii, and C. disporicum are found in both Budu. Nioella sediminis and S. baltica are found in Budu Pariaman. Peptostreptococcus russeli and C. novyi were found in Budu Pasaman. Conclusion: Metagenomic analysis of Budu from different fish, Pariaman (fresh fish) and Pasaman (frozen fish) showed that the biodiversity of bacteria was barely different. Both Budu found lactic acid bacteria from the Enterococcaceae family, genus Vagococcus, and pathogenic bacteria, such as S. cohnii, P. russeli, C. disporicum, and S. baltica. The discovery of various species of pathogenic bacteria indicates that development is still needed in the Budu production process to improve Budu quality. [J Adv Vet Anim Res 2023; 10(4.000): 801-808]

Objective: Various disease complications are a risk of overweight or obesity, so losing weight can reduce the risk of diseases caused by obesity. Binahong leaf ethanol extract (Anredera cordifolia) is a weight-loss herbal preparation. Aim: This study aims to analyze whether A. cordifolia extract is effective in losing weight by affecting the mechanism of adipogenesis in an animal obesity model. Materials and Methods: Animals were grouped into six groups as follows: the normal diet (K1), the negative control group (K2), the positive control group with Orlistat at a dose of 20 mg/kg BW (K3), an ethanol extract of A. cordifolia leaves at doses of 50 mg/kg BW (P1), 100 mg/kg BW group (P2), and 150 mg/kg BW (P3). All rats were fed a diet that consisted of high fat for eight weeks, except K1. Afterward, the treatments were given based on group distribution. Then, the rats were treated based on their groups for 4 weeks, and the high-fat diet was still given during the treatment for the control groups (K2). Anthropometric examinations such as body weight, length, and the circumference of the abdomen were measured. Metabolic parameters, including blood glucose, cholesterol levels, triglyceride levels, and abdominal fat weight, were measured using molecular parameters that measured PI3K levels and Extracellular signal-regulated kinase (ERK) in abdominal fat tissue samples using the ELISA method. Results: ERK levels of abdominal fat were lowered in the treatment group using the extract of A. cordifolia (50 mg/kg BW (P1) and 100 mg/kg BW (P2)) compared to the control group that was given a high-fat diet without treatment. The control group, which was fed a high-fat diet without treatment, had an average ERK level of 10.17 ± 2.98 ng/ml, P1 (50 mg/kg BW). Furthermore, when ethanol extracts were used as opposed to the control group, which received a high-fat diet without treatment, there was an increase in phosphoinositide three-kinase (PI3K) levels (K2). The control group received 9.35 ± 2.87 ng/ml, the treatment group received 100 mg/kg BW (P2) 9.48 ± 1.54 ng/ml, and the treatment group received 150 mg/kg BW (P3) 7.87 ± 1.79 ng/ml. The weight of fat in the abdomen differed between the groups that received a high-fat diet without treatment (K2) and those that received a high-fat diet with treatment (P1, P2, P3; p < 0.05). Conclusion: Anredera cordifolia extract possesses anti-obesity activities by decreasing ERK and increasing PI3K levels, as well as reducing abdominal fat weight. J. Adv. Vet. Anim. Res., 10(4): 809–816, December 2023 http://doi.org/10.5455/javar.2023.j737

Objective: Various disease complications are a risk of overweight or obesity, so losing weight can reduce the risk of diseases caused by obesity. Binahong leaf ethanol extract (Anredera cordifolia) is a weight-loss herbal preparation. Aim: This study aims to analyze whether A. cordifolia extract is effective in losing weight by affecting the mechanism of adipogenesis in an animal obesity model. Materials and Methods: Animals were grouped into six groups as follows: the normal diet (K1), the negative control group (K2), the positive control group with Orlistat at a dose of 20 mg/kg BW (K3), an ethanol extract of A. cordifolia leaves at doses of 50 mg/kg BW (P1), 100 mg/kg BW group (P2), and 150 mg/kg BW (P3). All rats were fed a diet that consisted of high fat for eight weeks, except K1. Afterward, the treatments were given based on group distribution. Then, the rats were treated based on their groups for 4 weeks, and the high-fat diet was still given during the treatment for the control groups (K2). Anthropometric examinations such as body weight, length, and the circumference of the abdomen were measured. Metabolic parameters, including blood glucose, cholesterol levels, triglyceride levels, and abdominal fat weight, were measured using molecular parameters that measured PI3K levels and Extracellular signal-regulated kinase (ERK) in abdominal fat tissue samples using the ELISA method. Results: ERK levels of abdominal fat were lowered in the treatment group using the extract of A. cordifolia (50 mg/kg BW (P1) and 100 mg/kg BW (P2)) compared to the control group that was given a high-fat diet without treatment. The control group, which was fed a high-fat diet without treatment, had an average ERK level of 10.17 ± 2.98 ng/ml, P1 (50 mg/kg BW). Furthermore, when ethanol extracts were used as opposed to the control group, which received a high-fat diet without treatment, there was an increase in phosphoinositide three-kinase (PI3K) levels (K2). The control group received 9.35 ± 2.87 ng/ml, the treatment group received 100 mg/kg BW (P2) 9.48 ± 1.54 ng/ml, and the treatment group received 150 mg/kg BW (P3) 7.87 ± 1.79 ng/ml. The weight of fat in the abdomen differed between the groups that received a high-fat diet without treatment (K2) and those that received a high-fat diet with treatment (P1, P2, P3; p < 0.05). Conclusion: Anredera cordifolia extract possesses anti-obesity activities by decreasing ERK and increasing PI3K levels, as well as reducing abdominal fat weight. [J Adv Vet Anim Res 2023; 10(4.000): 809-816]

Heartwater, one of the major tick-borne diseases of some domestic and wild ruminants in Africa, is caused by Ehrlichia ruminantium. The genetic diversity of E. ruminantium isolates renders the available vaccine ineffective against certain virulent isolates. To better understand the E. ruminantium genotypes in South Africa, a total of 1004 Amblyomma hebraeum tick deoxyribonucleic acid (DNA) samples from cattle in three South African provinces were tested by pCS20 Sol1 real-time polymerase chain reaction (qPCR) and characterised by multilocus sequence typing (MLST) using five housekeeping genes. Out of 1004 samples tested, 222 (22%) were positive for E. ruminantium. The occurrence of E. ruminantium in Mpumalanga, KwaZulu-Natal and Limpopo provinces was 19%, 22% and 27%, respectively. The E. ruminantium positive samples were screened for housekeeping genes and sequenced. Phylogenetic analysis revealed three main lineages: clade 1 made up of worldwide isolates (eastern, southern Africa, and Caribbean isolates), clade 2 comprised only West African isolates and clade 3 consisted of Omatjenne, Kümm2 and Riverside. Some study sample sequences were not identical to any of the reference isolates. However, they could all be grouped into the worldwide clade. Genetic variation in the sequenced regions was observed in the form of single nucleotide polymorphisms (SNPs). Using MLST to characterise E. ruminantium field isolates allowed the South African genotypes to be clearly distinguished from the distinct West African isolates.Contribution: Characterisation of E. ruminantium field isolates is important for the control of heartwater and contributes to preliminary knowledge required for the development of a more practical vaccine against heartwater.

Heartwater, one of the major tick-borne diseases of some domestic and wild ruminants in Africa, is caused by Ehrlichia ruminantium. The genetic diversity of E. ruminantium isolates renders the available vaccine ineffective against certain virulent isolates. To better understand the E. ruminantium genotypes in South Africa, a total of 1004 Amblyomma hebraeum tick deoxyribonucleic acid (DNA) samples from cattle in three South African provinces were tested by pCS20 Sol1 real-time polymerase chain reaction (qPCR) and characterised by multilocus sequence typing (MLST) using five housekeeping genes. Out of 1004 samples tested, 222 (22%) were positive for E. ruminantium. The occurrence of E. ruminantium in Mpumalanga, KwaZulu-Natal and Limpopo provinces was 19%, 22% and 27%, respectively. The E. ruminantium positive samples were screened for housekeeping genes and sequenced. Phylogenetic analysis revealed three main lineages: clade 1 made up of worldwide isolates (eastern, southern Africa, and Caribbean isolates), clade 2 comprised only West African isolates and clade 3 consisted of Omatjenne, Kümm2 and Riverside. Some study sample sequences were not identical to any of the reference isolates. However, they could all be grouped into the worldwide clade. Genetic variation in the sequenced regions was observed in the form of single nucleotide polymorphisms (SNPs). Using MLST to characterise E. ruminantium field isolates allowed the South African genotypes to be clearly distinguished from the distinct West African isolates. Contribution: Characterisation of E. ruminantium field isolates is important for the control of heartwater and contributes to preliminary knowledge required for the development of a more practical vaccine against heartwater.

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.

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.

Clinical signs suggestive of peste des petits ruminants (PPR) involved herds of small ruminants, which were described elsewhere in Sudan. Peste des petits ruminants was confirmed using an Immunocapture ELISA (IC-ELISA) assay in samples of infected and dead animals in areas of outbreaks. Therefore, to update information regarding the current situation and for assessment of the serological prevalence of PPR in small ruminants mingled at Central and Western Sudan during 2018–2019, 368 sera were collected from sheep (325 sera) and goats (43 sera) with different ages and breeds. These sera included 186 sera (173 sheep and 13 goats) from White Nile State and 182 sera (152 sheep and 30 goats) from Kordofan States. Competitive ELISA demonstrated higher prevalence of PPRV antibodies of 88.9%, 90.7% and 88.6% in both sheep and goats, goats, and sheep sera, respectively. Moreover, 100%, 94.7% and 78.5% seroprevalence values were demonstrated in South Kordofan, North Kordofan and White Nile States. The higher seroprevalence values detected in sera of unvaccinated sheep and goats indicated the wide exposure of these animals to PPRV and presence of protection following PPR viral infection. The findings of the study indicated that PPR is endemic in the surveyed areas of Sudan.Contribution: The study will contribute effectively to the global eradication programme of PPR organised by the World Organization for Animal Health (WOAH, formerly OIE) and Food and Agriculture Organization (FAO). To completely eliminate PPR from Sudan by 2030, local efforts should be directed towards effectively and wholly vaccinating small ruminants using PPRV vaccine especially in routes of seasonal animal’s movement and shared grazing areas.

Clinical signs suggestive of peste des petits ruminants (PPR) involved herds of small ruminants, which were described elsewhere in Sudan. Peste des petits ruminants was confirmed using an Immunocapture ELISA (IC-ELISA) assay in samples of infected and dead animals in areas of outbreaks. Therefore, to update information regarding the current situation and for assessment of the serological prevalence of PPR in small ruminants mingled at Central and Western Sudan during 2018–2019, 368 sera were collected from sheep (325 sera) and goats (43 sera) with different ages and breeds. These sera included 186 sera (173 sheep and 13 goats) from White Nile State and 182 sera (152 sheep and 30 goats) from Kordofan States. Competitive ELISA demonstrated higher prevalence of PPRV antibodies of 88.9%, 90.7% and 88.6% in both sheep and goats, goats, and sheep sera, respectively. Moreover, 100%, 94.7% and 78.5% seroprevalence values were demonstrated in South Kordofan, North Kordofan and White Nile States. The higher seroprevalence values detected in sera of unvaccinated sheep and goats indicated the wide exposure of these animals to PPRV and presence of protection following PPR viral infection. The findings of the study indicated that PPR is endemic in the surveyed areas of Sudan. Contribution: The study will contribute effectively to the global eradication programme of PPR organised by the World Organization for Animal Health (WOAH, formerly OIE) and Food and Agriculture Organization (FAO). To completely eliminate PPR from Sudan by 2030, local efforts should be directed towards effectively and wholly vaccinating small ruminants using PPRV vaccine especially in routes of seasonal animal’s movement and shared grazing areas.

Lymnaea natalensis is the only snail intermediate host of Fasciola gigantica, the causative agent of fascioliasis, in Nigeria. The species also serves as intermediate host for many other African trematode species of medical and veterinary importance, and it is found throughout the country. However, there is no detailed information on the factors that influence its distribution and seasonal abundance in the tropical aquatic habitats in Nigeria. This study used the geographic information system and remotely sensed data to develop models for predicting the distribution of L. natalensis in South-Western Nigeria. Both land surface temperature (LST) and normalised difference vegetation index (NDVI) were extracted from Landsat satellite imagery; other variables (slope and elevation) were extracted from a digital elevation model (DEM) while rainfall data were retrieved from the European Meteorology Research Programme (EMRP). These environmental variables were integrated into a geographic information system (GIS) to predict suitable habitats of L. natalensis using exploratory regression. A total of 1410 L. natalensis snails were collected vis-à-vis 22 sampling sites. Built-up areas recorded more L. natalensis compared with farmlands. There was no significant difference in the abundance of snails with season (p  0.05). The regression models showed that rainfall, NDVI, and slope were predictors of L. natalensis distribution. The habitats suitable for L. natalensis were central areas, while areas to the north and south were not suitable for L. natalensis.Contribution: The predictive risk models of L. natalensis in the study will be useful in mapping other areas where the snail sampling could not be conducted.

Lymnaea natalensis is the only snail intermediate host of Fasciola gigantica, the causative agent of fascioliasis, in Nigeria. The species also serves as intermediate host for many other African trematode species of medical and veterinary importance, and it is found throughout the country. However, there is no detailed information on the factors that influence its distribution and seasonal abundance in the tropical aquatic habitats in Nigeria. This study used the geographic information system and remotely sensed data to develop models for predicting the distribution of L. natalensis in South-Western Nigeria. Both land surface temperature (LST) and normalised difference vegetation index (NDVI) were extracted from Landsat satellite imagery; other variables (slope and elevation) were extracted from a digital elevation model (DEM) while rainfall data were retrieved from the European Meteorology Research Programme (EMRP). These environmental variables were integrated into a geographic information system (GIS) to predict suitable habitats of L. natalensis using exploratory regression. A total of 1410 L. natalensis snails were collected vis-à-vis 22 sampling sites. Built-up areas recorded more L. natalensis compared with farmlands. There was no significant difference in the abundance of snails with season (p  0.05). The regression models showed that rainfall, NDVI, and slope were predictors of L. natalensis distribution. The habitats suitable for L. natalensis were central areas, while areas to the north and south were not suitable for L. natalensis. Contribution: The predictive risk models of L. natalensis in the study will be useful in mapping other areas where the snail sampling could not be conducted.

Brucellosis is a major threat to public health especially in developing countries including Pakistan. This study reveals the characterisation of Brucella species affecting humans and goats in the Swat region of Khyber Pakhtunkhwa, Pakistan. Blood samples were collected from shepherds and goats and analysed by Rose Bengal precipitation test (RBPT), standard plate agglutination test (SPAT), polymerase chain reaction (PCR) and Sanger sequencing of 16S rRNA gene. The findings of the study indicated 24% (36/150) and 11.3% (17/150) positivity for Brucella abortus and Brucella melitensis, respectively, in human samples. In samples of goats, 26.66% (40/150) were positive for B. abortus and 16.66% (25/150) samples were positive B. melitensis by SPAT. The species-specific PCR confirmed B. abortus in 24% (36/150) of human samples and 26.66% (17/150) of goat samples by targeting the IS711 locus. The remaining seropositive samples were confirmed as B. melitensis using IS711 M species-specific primer. The sequences of the amplified fragments of the 16S rRNA gene were blasted, and phylogenetic analysis revealed that Brucella species circulating in the Swat district were closely related to B. melitensis and B. abortus reported from India, China, Philippines, and the United States (US) showing the existence of the possible epidemiological linkage among the Brucella species. This study concluded that there was a higher prevalence of B. abortus (26.6%) in humans and goats compared to B. melitensis (16.6%). These results revealed that the Brucella species were circulating in both humans and goats in the study areas. The findings of the study concluded that B. abortus and B. melitensis were circulating in goats and shepherds with a higher prevalence of B. abortus than B. melitensis. Furthermore, the Brucella species identified in Swat were phylogenetically related to the Brucella species reported from India, China, Philippines and the US.Contribution: The proposed study covers the scope of the journal. The species of the genus Brucella affect both animals and shepherds. This study investigates the seroprevalence of brucellosis in shepherds and goats in different geographical areas in the Swat district. The phylogenetic analysis of the Brucella spp. identified in Swat showed close relationships to the Brucella species reported in India, China, Philippines and the US, which shows the possible epidemiological linkages between the Brucella spp.

Brucellosis is a major threat to public health especially in developing countries including Pakistan. This study reveals the characterisation of Brucella species affecting humans and goats in the Swat region of Khyber Pakhtunkhwa, Pakistan. Blood samples were collected from shepherds and goats and analysed by Rose Bengal precipitation test (RBPT), standard plate agglutination test (SPAT), polymerase chain reaction (PCR) and Sanger sequencing of 16S rRNA gene. The findings of the study indicated 24% (36/150) and 11.3% (17/150) positivity for Brucella abortus and Brucella melitensis, respectively, in human samples. In samples of goats, 26.66% (40/150) were positive for B. abortus and 16.66% (25/150) samples were positive B. melitensis by SPAT. The species-specific PCR confirmed B. abortus in 24% (36/150) of human samples and 26.66% (17/150) of goat samples by targeting the IS711 locus. The remaining seropositive samples were confirmed as B. melitensis using IS711 M species-specific primer. The sequences of the amplified fragments of the 16S rRNA gene were blasted, and phylogenetic analysis revealed that Brucella species circulating in the Swat district were closely related to B. melitensis and B. abortus reported from India, China, Philippines, and the United States (US) showing the existence of the possible epidemiological linkage among the Brucella species. This study concluded that there was a higher prevalence of B. abortus (26.6%) in humans and goats compared to B. melitensis (16.6%). These results revealed that the Brucella species were circulating in both humans and goats in the study areas. The findings of the study concluded that B. abortus and B. melitensis were circulating in goats and shepherds with a higher prevalence of B. abortus than B. melitensis. Furthermore, the Brucella species identified in Swat were phylogenetically related to the Brucella species reported from India, China, Philippines and the US. Contribution: The proposed study covers the scope of the journal. The species of the genus Brucella affect both animals and shepherds. This study investigates the seroprevalence of brucellosis in shepherds and goats in different geographical areas in the Swat district. The phylogenetic analysis of the Brucella spp. identified in Swat showed close relationships to the Brucella species reported in India, China, Philippines and the US, which shows the possible epidemiological linkages between the Brucella spp.

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.

Objective: This study aimed to see if increasing the concentration of nutmeg flesh extract in vitro could increase the growth of Lactobacillus plantarum bacteria and if it had any effect on broiler chicken performance. Materials and Methods: Different concentrations of nutmeg flesh extract (5, 10, 15, and 20/100 ml distilled water) were combined with 10 ml L. plantarum (bacterial concentration 1 × 109 cfu/ ml) to produce synbiotics. A total of 250 unsexed Lohmann broiler chickens were reared together from 0 to 7 days of age in the in vivo study. Beginning on day 8, synbiotics nutmeg flesh extract and L. plantarum were added to the ration in amounts of 0.5, 1, 1.5, and 2 ml/kg for T1, T2, T3, and T4, respectively, while no synbiotics were added to the control diet (T0). Results: The levels of nutmeg flesh extract had a significant (p < 0.05) effect on L. plantarum growth. In the survival test against gastric acid, bile salts, and temperature, the addition of nutmeg flesh extract (20/100 ml distilled water) significantly (p < 0.05) maintained the population of L. plantarum. In vivo studies showed that the T1,T2,T3, and T4 groups gained more body weight (p < 0.05) than the T0 group during the rearing period but had no effect (p > 0.05) on the internal organ weight and carcass of broiler chickens. Conclusions: Nutmeg flesh extract could stimulate the growth of L. plantarum bacteria, and using it as a synbiotic could improve broiler chicken performance. J. Adv. Vet. Anim. Res., 10(1): 42–50, March 2023 http://doi.org/10.5455/javar.2023.j650

Objective: This study aimed to see if increasing the concentration of nutmeg flesh extract in vitro could increase the growth of Lactobacillus plantarum bacteria and if it had any effect on broiler chicken performance. Materials and Methods: Different concentrations of nutmeg flesh extract (5, 10, 15, and 20/100 ml distilled water) were combined with 10 ml L. plantarum (bacterial concentration 1 × 109 cfu/ ml) to produce synbiotics. A total of 250 unsexed Lohmann broiler chickens were reared together from 0 to 7 days of age in the in vivo study. Beginning on day 8, synbiotics nutmeg flesh extract and L. plantarum were added to the ration in amounts of 0.5, 1, 1.5, and 2 ml/kg for T1, T2, T3, and T4, respectively, while no synbiotics were added to the control diet (T0). Results: The levels of nutmeg flesh extract had a significant (p < 0.05) effect on L. plantarum growth. In the survival test against gastric acid, bile salts, and temperature, the addition of nut¬meg flesh extract (20/100 ml distilled water) significantly (p < 0.05) maintained the population of L. plantarum. In vivo studies showed that the T1,T2,T3, and T4 groups gained more body weight (p < 0.05) than the T0 group during the rearing period but had no effect (p > 0.05) on the internal organ weight and carcass of broiler chickens. Conclusions: Nutmeg flesh extract could stimulate the growth of L. plantarum bacteria, and using it as a synbiotic could improve broiler chicken performance. [J Adv Vet Anim Res 2023; 10(1.000): 42-50]