Molecular diagnosis of bovine tuberculosis on postmortem carcasses during routine meat inspection in Benin: GeneXpert® testing to improve diagnostic scheme.

Background and Aim: Bovine tuberculosis (TB) is a zoonotic disease of major public health importance, particularly in African countries, where control measures are limited or largely not applied. This study aimed to determine the accuracy of the currently used bovine TB diagnostic method at slaughterhouses in Benin; this is to contribute to the betterment and improvement in the epidemiological surveillance of the disease in the country. Materials and Methods: A total of 40 tissue samples were collected from meat/viscera (lung, liver, heart, kidney, and the gastro-intestinal tract tissues) at Cotonou slaughterhouses from ruminants suspected to be infected with bovine TB during routine meat inspection. The collected samples were analyzed using GeneXpert testing technique as a reference method. Results: Twenty-six samples tested positive out of the 40 suspected tissue samples collected by GeneXpert diagnostic technique; this shows the limitation of the routine meat inspection in detecting bovine TB as currently performed in Benin. Conclusion: The outcome of the use of the molecular technique, therefore, supports the importance of the use of a molecular tool alongside the routine meat inspection for a better understanding of the epidemiology of bovine TB in Benin. However, more robust technical and policy efforts are needed for a sustainable implementation of such a strategy.

Assessment of phenotypic diversity within Goliath chickens population in Benin.

Goliath chickens are a new strain of poultry that originated in the department of Zou, commune of Agbangnizoun in the Republic of Benin. The characteristics of this chicken strain have not been well studied. The purpose of this study was to characterize the phenotypic diversity of Goliath chickens in departments of Zou and Couffo. A total of 140 birds made up of 92 females and 48 males were sampled using 18 morphological descriptors. The results showed a wide variation in plumage colour within overall population with the most dominant being black (18.57%), white (17.14%) and red (16.43%). Black (21.74%) and white (20.65%) plumage dominated in females and red in males (35.42%). Coloured beaks and shanks were common in both chicken sexes. Yellow eyes were more common in females (63.04%), while red eyes were the most prevalent in males (43.75%). Comb and wattle colours were red in both sexes. Morphometrically, sexual dimorphism is very marked most of the body measurements with male birds being significantly superior. The average body weight was 3.26 kg in males and 2.78 kg in females. The population was clustered in three morphotypes according to the gradient size. Chickens from Bohicon and Agbangnizoun on the one hand, and those from Zagnanado and Toviklin on the other, formed groups 1 and 2, respectively, while group 3 included individuals from Djidja. 77.96% of observed diversity was due to variation between individuals. Molecular characterization is recommended for a thorough knowledge of the Goliath chicken strain of Benin.

Treatment of Girolando cattle dermatophilosis using a combination of different plant extracts in the municipality of Abomey-Calavi, Republic of Benin.

BACKGROUND AND AIM: Dermatophilosis is a bacterial infection of the skin of animals. It is prevalent worldwide and is caused by Dermatophilus congolensis. The study aimed to assess the therapeutic efficacy of different mixtures prepared with indigenous phytogenetic extracts from Benin in the management and treatment of Girolando cattle that showed high sensitivity to the disease compared to any other known cattle breed in Benin. MATERIALS AND METHODS: Consequently, two types of extact mixtures (extract mixture 1=Elaeis guineensis kernel oil + essential oil of Ocimum gratissimum + sap of Jatropha multifidi; extract mixture 2= O. gratissimum + sap extract of J. multifida) of 20% concentration were prepared and tested on eight Girolando cattle. Two different parts of the affected skin with acute lesions of dermatophilosis were debrided, and 0.5 mL of each of the extract mixture was applied per square centimeter of a single zone of the affected skin. RESULTS: Both extracts mixtures were found to possess significant wound healing properties compared to the control (procaine G penicillin). However, the extract mixture 1, which was made up of Elaeis guineensis kernel oil, essential oil of O. gratissimum and sap of J. multifida (Linn), showed a better result. This was evident by increase in the rate of wound contraction and healing without recurrence 2 weeks after the end of the experiment and the subsequent immediate manifestation of hair or hair growth at the affected area. CONCLUSION: The preliminary findings of this study are very promising. Extract mixture 1 could serve as an alternative in the treatment or management of bovine dermatophilosis in Benin and other dermatophilosis endemic areas of the world. However, in vitro testing and sensitivity against isolated D. congolensis organism using extract mixture 1 as well as cost implications should be studied.

Phenotypic and genotypic characterization of dog tick Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) resistance to deltamethrin in Benin.

The aim of this study was to evaluate the phenotypic and genotypic resistance of Rhipicephalus sanguineus sensu lato to deltamethrin in dogs in the municipality of Natitingou in Benin. In this study, the collection and identification of ticks, Larval Packet Test and specific allele PCR were the main methods used. A total of 41 dogs were examined and 420 ticks were collected in six areas which includes: Tchrimina, Yokossi, Boriyoure, Berécingou, Péporiyakou and Perma. Three species of ticks were identified and they include: R. sanguineus s.l., Rhipicephalus microplus and Haemaphysalis spp. However, R. sanguineus s.l. was the most abundant tick species (9.85) and had the highest parasitic intensity (10.36). The results of the Larval Packet Test showed that the larvae of R. sanguineus s.l. developed a phenotypic resistance against deltamethrin, particularly in the Bérécingou area where resistance ratios of LC50 and LC95 of R. sanguineus s.l. compared to a susceptible strain were 1.591 (1.025-3.054) and 65.339 (20.235-579.825) respectively. The molecular study of the sodium channel gene revealed three genotypes namely the susceptible genotype (SS), the heterozygous genotype (SR) and the resistant genotype (RR), which correlates with the phenotypic results. This information is fundamental to improve monitoring and resistance management strategies of R. sanguineus s.l. ticks to pyrethroids.

Assessment of the reproductive performances of native sows in Southern Benin.

Native sows contribute to a large extent to food security and poverty alleviation in Benin. However, their reproductive performance particularly under extensive systems is poorly characterized. The objective of this study was to fill this knowledge gap by selecting 284 multiparous sows based on hair color and some reproductive parameters. The results showed that native sows of Southern Benin can be clustered into three groups with group 3 showing the best reproductive performances including highest average litter size (LS, 10.31 piglets), live-born piglets (LBP, 10.31 piglets), number of functional teats (10.94), and shorter average farrowing interval (6 mo). The analysis of sows' performances based on their origin revealed that sows from agro-ecological zone 8 have the highest number of LBP and the lowest age at first mating and first farrowing. The analysis of sows' performances based on the hair color showed that those with black hair have the largest LS and the highest number of LBP. This study showed that Benin's native sows have good reproductive ability with enough variation to develop a sustainable pig industry for a better contribution to food security and wealth creation.

Inventory of biosecurity measures and antibiotics therapy practices on laying hen farms in Benin.

BACKGROUND AND AIM: Laying hen breeding is on the rise in Benin; nevertheless, there are several sanitary constraints to its development, including bacterial diseases. Faced with this situation, breeders mainly resort to different means of treatment. The objective of this study was to assess the current state of hygiene measures, the bacterial diseases commonly encountered, and antibiotic therapy practices on laying hen farms in Benin. MATERIALS AND METHODS: A total of 200 laying hen farms were randomly selected from lists of laying hen farms obtained from veterinary offices, territorial agricultural development agencies, and the Benin National Union of Professional Aviculturists. Each visited farmer was subjected to a semi-structured questionnaire by direct interview. The results were compared using the bilateral Z-test. RESULTS: The results of this survey revealed that 99.5% of the surveyed farms had a health and medical prophylaxis program although only 88.5% of them reported strictly adhering to it (p<0.001). About 25.0% of them reported that the dominant bacterial diseases they commonly encountered on their farms were salmonellosis, colibacillosis, and chronic respiratory disease. Only 7.0% of farmers said that they confirmed their diagnosis outside of clinical signs through laboratory analysis. To control these pathologies, 14.5% of farmers used only oxytetracycline, while 39.0% used other antibiotics such as colistin, enrofloxacin, tylosin, tylodox, flumequine, and norfloxacin. In comparison, 13.5% used a trimethoprim-sulfadimethoxine and sulfadimidine combination, while 32.0% said that they used erythromycin, oxytetracycline, streptomycin, neomycin, and colistin (p<0.001) combination. CONCLUSION: This study highlights the inadequacies of hygiene and antibiotic therapy practices implemented on Benin's laying hen farms.

Preliminary study on the tick population of Benin wildlife at the moment of its invasion by the Rhipicephalus microplus tick (Canestrini, 1888).

BACKGROUND AND AIM: Rhipicephalus microplus (Rm) is one of the most problematic livestock tick species in the world. Its rapid propagation and resistance to acaricides make it control difficult in the sub-region and Benin particularly. The aim of this work was to check its presence in wildlife and to confirm the possible role of reservoir wildlife may play in the propagation of the parasite. This will help to design more efficient control strategy. MATERIALS AND METHODS: This study was conducted from February to March 2017 in the National Parks of Benin (Pendjari and W Park) and wildfowl's assembly and selling point in Benin. Ticks were manually picked with forceps from each animal after slaughtering by hunters then stored in 70° ethanol. Collected ticks were counted and identified in the laboratory using the identification key as described by Walker. RESULTS: Overall, seven species of ticks (Amblyomma variegatum, Boophilus decoloratus, Rm, Boophilus spp., Hyalomma spp., Rhipicephalus sanguineus, Rhipicephalus spp.) were identified on nine wild animal species sampled (Cane rat, wildcat, Hare, Doe, Cricetoma, Buffalo, Buffon Cobe, and Bushbuck and Warthog). The average number of ticks varies from 3 to 6 between animal species, 3 to 7 between localities visited, and 2 to 5 between tick species. However, these differences are statistically significant only for localities. Considering tick species and animal species, the parasite load of Rm and Rhipicephalus spp. is higher; the buffalo being more infested. The analysis of deviance reveals that the abundance of ticks observed depends only on the observed localities (p>0.05). However, the interactions between animal species and localities on the one hand and between animal and tick species on the other hand, although not significant, have influenced the abundance of ticks as they reduce the residual deviance after their inclusion in the model. CONCLUSIONS: This study reported the presence of Rm in wildlife of Benin and confirmed its role in the maintenance and spread of the parasites. It is, therefore, an important risk factor that we must not neglect in the epidemiological surveillance and ticks control strategies in the West African sub-region and particularly in Benin.

Sensitivity of Different Cattle Breeds to the Infestation of Cattle Ticks Amblyomma variegatum, Rhipicephalus microplus, and Hyalomma spp. on the Natural Pastures of Opkara Farm, Benin.

A study was carried out on the Opkara (Benin) cattle farm on 64 cattle of four different breeds (16 individuals per breed) from June to December 2016. During this study, three tick species were found in different numbers, Amblyomma variegatum (732), Rhipicephalus microplus (8079), and Hyalomma spp. (208), with parasitic intensity of 11.90, 126.23, and 3.25, respectively. The interracial comparison of the tick infestation between the cattle showed a significant difference (P < 0.001). However, Girolando was more infested than all the cattle breeds. Infestation of A. variegatum, R. microplus, and Hyalomma spp. on the Girolando was, respectively, 19.43 ± 2.71, 171.25 ± 23.50, and 7.12 ± 0.63, but the Borgou were less infested. Borgou breed females were more infested by A. variegatum (4.41 ± 1.14) than females Girolando (4.20 ± 0.90). The Crossbred and Azawak females were less infested (P < 0.01). The mean of A. variegatum on Borgou, Azawak, Crossbred, and Girolando calves was 1.29 ± 0.35, 0.66 ± 0.26, 1.37 ± 0.37, and 2.25 ± 0.48 (P < 0.01), respectively. The results of this study can be exploited to include genetic and nongenetic approaches to tick control.

West African Cattle Farmers' Perception of Tick-Borne Diseases.

Worldwide, cattle production is struggling to face the negative impacts caused by ticks and Rhipicephalus (Boophilus) microplus is one of the most harmful ticks for livestock. Most of the people in West Africa depend on cattle farming and subsistence agriculture. The presence of ticks on cattle is a major problem faced by smallholder farmers who fight for their livelihood. National and regional tick control programs could assist these rural communities in protecting their livelihoods against ticks and tick-borne diseases, but only if they take into account the targeted herders and their perception on cattle management and tick control. This paper aims to provide a better insight in the socio-economic characteristics of Beninese cattle farmers, and their perception on tick burden, as well as to document common tick control strategies. Different tick species and their seasonality are well understood by cattle herders. For tick control, many still use manual tick removal, especially in the north of the country. The high cost of acaricides, the lack of financial means of African farmers, and of the local stockbreeders in particular, limits the use of acaricides in livestock breeding in Benin. While aiming to increase the meat or milk production of their animals, stockbreeders who can afford it sometimes turn to an abusive use of acaricides, which might in time lead to an increase in tick resistance. This study remains one of the rare studies to report extensively on the perceptions of West African cattle herders.

Evidence of multiple mechanisms of alphacypermethrin and deltamethrin resistance in ticks Rhipicephalus microplus in Benin, West Africa.

Ticks are obligate haematophagous arthropods, causing heavy losses in affected livestock. The objective of this study is to investigate phenotypic and genotypic resistance in Rhipicephalus microplus populations from Benin. Engorged female adult ticks were collected from cattle in two districts of Benin. Bioassays, biochemical and molecular tests were carried out on these ticks to determine the phenotypic, enzymatic and genetic status of resistance. Results of bioassays showed high resistance factors (RF > 41). The molecular tests showing the presence of the domain II mutation and absence of the domain III mutation in the voltage-gated sodium channel gene. Biochemical tests showed increased activity of esterases, multifunction oxidases and glutathione transferases in resistant samples. Genotyping the samples showed high levels of heterozygous genotypes (73.36% and 63.30%) as compared to homozygous susceptible and resistant genotypes (23.3% and 10%) respective at Samiondji and Betecoucou. A correlation between phenotype resistance and presence of the domain II mutation at the voltage gated sodium channel gene was observed suggesting that this could be associated with resistance. Target site mutation and metabolic detoxification are mechanisms of resistance to pyrethroids in R. microplus tick populations from Benin.

Toxoplasma gondii infection in meat animals from Africa: Systematic review and meta-analysis of sero-epidemiological studies.

AIM: Toxoplasma gondii is an ubiquitous apicomplexan parasite which causes toxoplasmosis in humans and animals. Felids especially cats are definitive hosts and almost all warm-blooded mammals, including livestock and human can serve as intermediate hosts. Food animals can be reservoirs for T. gondii and act as one of the sources for parasite transmission to humans. The objective of this study is to collect serological data on the prevalence of anti-T. gondii antibody, and risk factors for certain food animals from Africa to provide a quantitative estimate of T. gondii infection among these species from different African countries. MATERIALS AND METHODS: Four databases were used to search seroepidemiological data on the prevalence of anti-T. gondii antibody in food animals between 1969 and 2016 from African countries. The search focused on data obtained by serologic test in food animals and meta-analyses were performed per species. RESULTS: A total of 30,742 individual samples from 24 countries, described in 68 articles were studied. The overall estimated prevalence for toxoplasmosis in chicken, camel, cattle, sheep, goat, pig were 37.4% (29.2-46.0%), 36% (18-56%), 12% (8-17%), 26.1% (17.0-37.0%), 22.9% (12.3-36.0%), and 26.0% (20-32.0%), respectively. Moreover, major risk factor of infection was age, farming system, and farm location. CONCLUSIONS: A significant variation in the seroepidemiological data was observed within each species and country. The results can aid in an updated epidemiological analysis but also can be used as an important input in quantitative microbial risk assessment models. Further studies are required for a better and continual evaluation of the occurrence of this zoonotic infection.

An Optimized Trichloroacetic Acid/Acetone Precipitation Method for Two-Dimensional Gel Electrophoresis Analysis of Qinchuan Cattle Longissimus Dorsi Muscle Containing High Proportion of Marbling.

Longissimus dorsi muscle (LD) proteomics provides a novel opportunity to reveal the molecular mechanism behind intramuscular fat deposition. Unfortunately, the vast amounts of lipids and nucleic acids in this tissue hampered LD proteomics analysis. Trichloroacetic acid (TCA)/acetone precipitation is a widely used method to remove contaminants from protein samples. However, the high speed centrifugation employed in this method produces hard precipitates, which restrict contaminant elimination and protein re-dissolution. To address the problem, the centrifugation precipitates were first grinded with a glass tissue grinder and then washed with 90% acetone (TCA/acetone-G-W) in the present study. According to our result, the treatment for solid precipitate facilitated non-protein contaminant removal and protein re-dissolution, ultimately improving two-dimensional gel electrophoresis (2-DE) analysis. Additionally, we also evaluated the effect of sample drying on 2-DE profile as well as protein yield. It was found that 30 min air-drying did not result in significant protein loss, but reduced horizontal streaking and smearing on 2-DE gel compared to 10 min. In summary, we developed an optimized TCA/acetone precipitation method for protein extraction of LD, in which the modifications improved the effectiveness of TCA/acetone method.