Identification and validation of novel breed-specific biomarker for the purpose of village chicken authentication using genomics approaches.

Local village chicken, or "Ayam kampung" as it's known in Malaysia, is considered a premium chicken breed with a higher price than other chicken breeds. As a result of their comparable appearances and sizes, colored broiler chickens are often sold as village chickens, which is a form of food fraud that can result in a 3- to 4-fold rise in profit. Therefore, developing a breed-specific authentication method is crucial for preventing food fraud in the poultry industry. This study aims to investigate the genetic diversity of village chickens from other commercial chicken breed populations available in the market (broiler [Cobb], colored broiler [Hubbard], and layer [DeKalb]) to identify breed-specific DNA fragments as biomarkers for village chicken authentication. The Whole-genome sequencing and mutation calling of 12 chickens (3 chickens/breed) led to the identification of a total of 73,454,654 single nucleotide polymorphisms (SNP) and 8,762,338 insertion and deletions (InDel) variants, with more variants detected in the village chicken population (6,346,704 SNPs; 752,408 InDels) compared to commercial breeds. Therefore, this study revealed that village chickens were more genetically variable compared to other breeds in Malaysia. Furthermore, the breed-specific genomic region located on chromosome 1 (1:84,405,652) harboring SNP (C-T) with high discrimination power was discovered and validated which can be considered as a novel breed-specific biomarker to develop a method for accurate authentication of village chickens in Malaysia. This authentication method offers potentialw applications in the chicken industry and food safety.

Prevalence and Haplotypes of Toxoplasma gondii in Native Village Chickens and Pigs in Peninsular Malaysia.

Toxoplasma gondii is an important zoonotic foodborne parasite capable of infecting almost all warm-blooded animal species worldwide. Toxoplasmosis is usually acquired via ingestion of undercooked infected animal tissues resulting in life-threatening consequences for unborn foetus and immunocompromised individuals. A cross-sectional study was carried out to determine the prevalence of T. gondii infection, its associated risk factors in farms, and haplotypes isolated from the native village chicken and pig populations in Peninsular Malaysia. The seroprevalence of T. gondii in village chickens at the animal level was low at 7.6% (95% CI: 4.60-11.60), while at the farm level, it was 52.0% (95% CI: 31.30-72.20). For pigs, the animal-level seroprevalence of T. gondii was 3.0% (95% CI: 1.60-5.10), while the farm-level, it was 31.6% (95% CI: 12.60-56.60). The PCR-based DNA detection on meat samples from chickens (n = 250) and pork (n = 121) detected 14.0% (95% CI: 9.95-18.9) and 5.8% (95% CI: 2.4-11.6) positive, respectively. Six unique T. gondii haplotypes were isolated from the tissue samples. Multivariable logistic regression analysis showed that feeding the chickens farm-produced feeds and allowing wild animals access to pig farms were significant determinants for farm-level seropositivity. Providing hygienic and good quality feeds to chickens and increasing biosecurity in pig farms through prevention of access by wildlife may reduce the risk of transmission of T. gondii infection in the local chickens and pig farms.

The usefulness of maternally inherited genetic markers for phylogeographic studies in village chicken.

Phylogeography plays a major role in understanding micro and macroevolutionary processes dealing with evolutionary interpretations of geographical distribution. This field integrates information from molecular genetics, population genetics, demography, and phylogeny for the interpretation of the geographical distribution of lineages. The full mtDNA sequence and W chromosome polymorphisms were exploited to assess the usefulness of two maternally-inherited genetic markers for phylogeographic studies of village chickens. We studied 243 full mtDNA sequences from three countries (Iraq, n = 27; Ethiopia, n = 211; and Saudi Arabia, n = 5) and a 13-kb fragment of the W chromosome from 20 Iraqi and 137 Ethiopian female chickens. The results show a high level of genetic diversity for the mtDNA within and among countries as well as within populations. On the other hand, sequence analysis of the W chromosome shows low genetic diversity both within and among populations. Six full mtDNA haplogroups (A, B, C1, C2, D1, and E1) were observed and 25 distinct W haplotypes. The results support the effectiveness of full mtDNA sequences but not the W chromosome in tracing the maternal historical genome background with, however, weak within a country phylogeographic signal.

Epidemiological risk factors and phylogenetic affinities of Sarcocystis infecting village chickens and pigs in Peninsular Malaysia

@#The intake of food and water containing the Sarcocystis parasite has been linked to a number of outbreaks worldwide, including Malaysia. Nevertheless, the lack of surveys and epidemiological data on Sarcocystis infections in Malaysia makes it difficult to estimate its occurrence in humans and animals. A cross-sectional study was conducted to determine the prevalence of Sarcocystis and the risk factors associated with infection among village chickens and pigs reared under different farm managements in Peninsular Malaysia. Phylogenetic trees were constructed using partial fragments of the 18S rRNA gene and ITS1 sequences. In the present study, 680 sera samples were collected from village chickens (n=250) and commercial pigs (n=433) and anti-Sarcocystis antibodies were screened using the enzymelinked immunosorbent assays (ELISA) kit. At the animal level, the prevalence of Sarcocystis was 9.2% (95% CI: 5.92-13.48) and at the farm level, it was 64.0% (95% CI: 42.52-82.03) in village chickens. The animal-level seroprevalence of Sarcocystis for pigs was 3.7% (95% CI: 2.13-5.93) and 36.8% (95% CI: 16.29-61.64) at the farm-level. Polymerase Chain Reaction (PCR) was conducted on meat samples from various parts of village chickens (n=250) consisting of brain, heart, lung, and pectoralis muscle tissues, and pork (n=121) consisting of intercostal muscle, diaphragm, and tongue. Sarcocystis DNA was detected in 6.4% (95% CI: 4.60-11.60) of village chicken samples but zero in pork samples. A total of 11 unique Sarcocystis haplotypes were isolated from these tissue samples. Multivariable logistic regression analysis of the putative risk factors showed a statistically significant association between Sarcocystis infection in pigs and uncovered storage of feed. Although no zoonotic Sarcocystis was isolated in this study, we reported the first discovery of S. wenzeli in Malaysia.

Discerning risk and uncertainty with "better insurance" - How animal health interventions for village chickens influence farmers' perceptions on the prevention of poultry diseases.

High mortality in young chicks and deaths from Newcastle disease (ND) are the two major constraints to village chicken production in low-and middle-income countries. A 12-month intervention study was conducted across 174 households in two townships of Central Myanmar, to reduce these chicken mortalities. Interventions included ND vaccinations of village chickens and improved chick management. Following the Health Belief Model (HBM) framework, 81 village chicken farmers involved in the intervention study were repeatedly surveyed before and after the interventions to determine the level of change in farmers' knowledge about ND, perceptions about susceptibility and severity of ND virus infection, perceptions about benefits and barriers of ND vaccination, as well as farmers' actions to improve the health status of their birds and to improve biosecurity and ND control measures. Marginal homogeneity tests were used to compare the level of change in farmers' matched responses to individual survey questions before and after the interventions. In addition, confirmatory factor analysis and structural equation modelling was used to create latent variables describing farmers' perceptions about susceptibility and severity of ND in their birds, farmers' perceptions about the barriers and benefits of conducting ND vaccination as well as farmers' cues to action and self-efficacy before and after the interventions. Interventions resulted in 33% more chicks surviving, which resulted in increased sales of birds after they had reached a market age of 7 months. After the interventions, farmers acknowledged the value of protecting birds from ND to increase their income (p = 0.0002). Farmers were also more willing to overcome barriers to vaccination (i.e., 'catching' birds for ND vaccinations) after the interventions (p = 0.0388). Interestingly, after the interventions, fewer farmers were concerned about the risk of their birds becoming infected with ND virus through cock fights and non-confinement (p = 0.0008 and p = 0.00022, respectively), and farmers were more uncertain about the susceptibility of their birds to ND virus infection after the interventions. Surprisingly, farmers became 'less active' in ensuring good biosecurity practices after the interventions: fewer farmers were willing to observe disease signs in village chickens (p < 0.001) and fewer farmers buried dead birds (p < 0.001). Perceived susceptibility was the only HBM component that was significantly reduced (p = 0.014) after the interventions: thus, less farmers were concerned about ND even when using management practices associated with high risk of ND transmission (e.g., cook fights, no confinement). Our results highlight the importance of information campaigns and extension work to ensure farmers maintain a high level of biosecurity when animal health interventions are implemented.

Chicken Authentication and Discrimination via Live Weight, Body Size, Carcass Traits, and Breast Muscle Fat Content Clustering as Affected by Breed and Sex Varieties in Malaysia.

Nowadays, the high demand for village chickens in Malaysia leads to the fraudulent substitution of indigenous chickens with other cheaper counterparts. Discriminating different chicken breeds based on their phenotypic characteristics is one strategy to avoid chicken adulteration. The main objective of this study was to authenticate and group dominant chicken breeds in Malaysia, including commercial chickens (Cobb, Hubbard, DeKalb) and cross-bred village chickens (Ayam Kampung, Akar Putra). The further discrimination of village chickens from underaged colored broilers (UCBs) (Hubbard, Sasso) was performed based on phenotype traits. The results showed that the breed had a significant effect (p < 0.05) on phenotypic characteristics, while the sex effect was not significant for some characteristics. In the first phase, the most remarkable discriminating factors were abdominal fat weight, breast muscle weight, chest circumference, shank length, and wingspan. However, in the second phase, notable variations in phenotypic characteristics between village chickens and UCBs were not detected. Principal component analysis (PCA) showed the successful separation of village chickens from high-performance breeds (broiler and colored broiler). Nevertheless, there was overlap among observations for Sasso and village chickens, which approved the possible similarities in their phenotypic characteristics. This study showed clear breed clustering, which leads to the chicken authentication based on their phenotypic characteristics.

Fatty acid profile in the breast and thigh muscles of the slow- and fast-growing birds under the same management system.

The aim of the study was to assess the effect of feeding the same diet to different breeds of chickens and at different ages on fatty acid (FA) composition of the breast and thigh muscles. A total of 150 chickens comprising 50 each of red jungle fowl (RJ) and village chicken (VC), the slow-growing birds, and the commercial broiler (CB), fast-growing birds, were used for this study. Ten chickens from each breed were serially euthanized at days 1, 10, 20, 56, and 120 post hatch, and pectoralis major and bicep femoris were harvested to represent the breast and thigh muscles respectively. It was revealed that the breast muscle concentrations of saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), and total polyunsaturated fatty acids (PUFA) are significantly different (p<0.05) among the breeds. Also, the FA composition of breast and thigh muscles among RJ, VC, and CB at various ages studied varied significantly (p<0.05) but without a definite pattern. The composition of MUFA was lower, but that of PUFA was higher in the RJ and VC than in the CB breast muscles. Within the breeds, the composition of total MUFA decreased, while that of PUFA increased with age. The total MUFA and PUFA showed no significant difference (p>0.05) between the breast and thigh muscles at different ages evaluated. This study suggests that slow-growing birds (RJ and VC) might be better sources of desirable FA than the fast-growing birds, CB.

Research Note: Molecular surveillance of Avian Paramyxovirus type-1 in nonvaccinated village chickens in Central Rift Valley of Oromia, Ethiopia.

Newcastle disease (ND) is a major infectious disease of poultry caused by a virulent strain of Avian Paramyxovirus type-1 (APMV-1). It is a major threat to the poultry industry in many countries of the world including Ethiopia. The aim of this study was to conduct molecular surveillance of ND Virus and identify potential risk factors for nonvaccinated village chicken in Central Rift Valley of Oromia, Ethiopia. A total of 84 pooled swab samples, each made from pools of 5 swabs for analysis, from cloacal and tracheal sites of chickens in the Central Rift Valley were collected, and RNA was extracted to carry out real-time quantitative polymerase chain reaction. Out of the 84 pooled swab samples tested for M-gene, 13 (15.48%) samples were found positive for APMV-1. The prevalence of ND in males was found to be 16.10% and that in females was 14.67%. Although the overall ND prevalence was 15.48% (13/84), the highest prevalence was recorded in Adama, 42.86% (6/14), and no positive case was observed in Bote and Bishoftu (P < 0.05), while intermediate prevalence was obtained from Batu, Arsi-negele, and Shashemene (P > 0.05). In general, the present study provides important information on the epidemiology of ND based on M-gene assay in Central Rift Valley of Oromia, Ethiopia, and highlights the importance of implementing molecular surveillances practice in live poultry markets and village chickens.

Participatory assessment of paid vaccination campaigns for village chickens against Newcastle disease in Kongo Central province.

In the Democratic Republic of Congo (DRC), where state-driven animal vaccination campaigns are organized only in response to epidemics, the organization of a permanent animal vaccination service is urgently needed. A non-governmental organization has set up an experimental paid vaccination service for village chickens against Newcastle Disease (ND) in the Kongo Central province. This mixed-method study presents a participatory assessment of this experiment, identifying socio-economic factors that influence the decision of chicken keepers to adopt vaccination. The study was conducted in four territories of the province. Qualitative and quantitative data were collected through 12 focus group discussions (FGDs) with professionals of animal health and chicken keepers and 160 semi-structured interviews with chicken keepers, sampled by snowball technique. This participatory process has resulted in the design of a grid for assessing animal vaccination service's performance. Here translating the narratives into a preliminary structured assessment, this grid is an output of the study, to be mobilized for future rapid assessments of the vaccination service in a quantitative prospect. The grid consisted of nine criteria, further composed by 16 items, translated into questions to be asked to chicken keepers and vaccinators. In our study area, fieldworkers enumerated four animal vaccination campaigns during a period of 21 years (except those subject to the present assessment). Around 13% of chicken keepers of our sample had participated in ND vaccination programs. Almost 96% of interviewed chicken keepers expressed their willingness to pay for ND vaccination, and 87% of chicken keepers that vaccinated their chickens perceived the vaccine as effective. Vaccinators estimated that 56% of the chicken keepers who were contacted had actually paid for the vaccination of their chickens. The assessment grid highlighted four points in favor of the sustainability of this service, i.e. the general interest of chickens keepers, vaccine efficacy, vaccine availability and ease of use of the vaccine. Two weak points were identified, viz. the poor access of chicken keepers to information and the weak motivation of vaccinators. The vaccine coverage was calculated within the sample at 13.1%. Paid vaccination campaign for village chicken in Kongo Central obtained a performance score of 62.8%, with the highest score in Kwilu-Ngongo (73.1%) and the lowest in Kasangulu (52.4%). Two factors of adoption of vaccination were identified as statistically significant, i.e. chicken housing and territory. Significant differences appeared between territories in access to information for chicken keepers and in vaccinators motivation. The priorities for the improvement of this service appear to be awareness raising among chicken keepers and increasing vaccinators' motivation.

Salmonella in native "village" chickens (Gallus domesticus): prevalence and risk factors from farms in South-Central Peninsular Malaysia.

Village chicken or Ayam Kampung, common to Southeast Asian countries, has always been regarded as superior in comparison to commercial broiler chicken in terms of wholesomeness and health benefits. The current study investigates the prevalence and risk factors of Salmonella among village chicken flocks from the central and southern states of Peninsular Malaysia. A total of 35 village flocks were sampled from Selangor (n = 19), Melaka (n = 10), Johor (n = 4), and Negeri Sembilan (n = 2). In total, 1,042 samples were collected; these included cloacal swabs (n = 675), eggs (n = 62), pooled drinking water (n = 175), pooled feeds (n = 70), and pooled flies (n = 60). Isolation of Salmonella from cloacal swabs, poultry drinking water, and feeds was carried out according to the protocols and recommendations of the World Organization for Animal Health (OIE) terrestrial manual. The prevalence of Salmonella at an individual bird-level was 2.5% (17/675, 95% CI: 1.6 to 4.0). All eggs screened were negative; in the case of environmental samples, however, Salmonella was detected in 5.14% (9/175), 7.14% (5/70), and 5.0% (3/60) for water, feed, and flies, respectively. A total of 34 isolates and 8 Salmonella serotypes were identified. Weltevreden (20.6%) was the most common, followed by Typhimurium and Agona (17.6%), Albany and Enteritidis (8.8%), Molade (5.9%), Corvallis and Schleissheim (2.9%), and others grouped as Salmonella spp. (11.8%). Multivariable logistic regression models revealed that Salmonella positivity among flocks could be strongly predicted by storage of feeds (uncovered feeds; OR = 10.38; 95% CI: 1.25 to 86.39; p = 0.030) and uncovered water tanks (uncovered tank; OR = 6.43; 95% CI: 1.02 to 40.60; p = 0.048). The presence of Salmonella in village chickens in the study area was lower than that of commercial chickens in Malaysia.

Whole-Genome Resequencing of Red Junglefowl and Indigenous Village Chicken Reveal New Insights on the Genome Dynamics of the Species.

The red junglefowl Gallus gallus is the main progenitor of domestic chicken, the commonest livestock species, outnumbering humans by an approximate ratio of six to one. The genetic control for production traits have been well studied in commercial chicken, but the selection pressures underlying unique adaptation and production to local environments remain largely unknown in indigenous village chicken. Likewise, the genome regions under positive selection in the wild red junglefowl remain untapped. Here, using the pool heterozygosity approach, we analyzed indigenous village chicken populations from Ethiopia, Saudi Arabia, and Sri Lanka, alongside six red junglefowl, for signatures of positive selection across the autosomes. Two red junglefowl candidate selected regions were shared with all domestic chicken populations. Four candidates sweep regions, unique to and shared among all indigenous domestic chicken, were detected. Only one region includes annotated genes (TSHR and GTF2A1). Candidate regions that were unique to each domestic chicken population with functions relating to adaptation to temperature gradient, production, reproduction and immunity were identified. Our results provide new insights on the consequence of the selection pressures that followed domestication on the genome landscape of the domestic village chicken.

Rural household chicken management and challenges in the Upper River Region of the Gambia.

Systematic assessment and documentation of extensive livestock production systems are necessary to design or review breeding programs, extension services, and policies. This study therefore examined management practices and challenges concerning village chicken production in the Upper River Region of the Gambia. The data gathered with 45-variable semi-structured questionnaires were analyzed based on the household head's education status and gender. Illiteracy level was high, and only 38% indicated that they attended certain schools. There was a significant relationship (p < 0.05) and phi coefficient of 0.35 between household heads' education status and record keeping. None of the examined management practices had a statistically significant relationship (p > 0.05) with household heads' gender. Children (10-14 years) were mostly responsible for providing care to the chickens. Foundation and replacement stocks were mainly acquired through purchase (78%). Many households (74%) indicated supplementary feeding of their flocks but only 34% provided separate houses apart from the household dwellings. Newcastle (68%) was the most common disease. Sick birds were recognized by restlessness (34%) and diarrhea (28%). External parasites were mainly controlled through local practices (52%) while the most common way to dispose dead birds was to throw them away (88%). Up to 90% indicated no formal training on disease management and access to extension agents was also low (20%). Control of chicken movement was occasionally done to protect birds from predators (60%) and then, to avoid contagious diseases (38%). Individual farmers and their associations need to be supported by stakeholders to access relevant information and uptake improved management techniques.

Natural resistance of Sri Lankan village chicken to Salmonella gallinarum infection.

1. An experiment was conducted to compare the natural resistance of an indigenous breed of local village chickens to Salmonella gallinarum with two commercial breeds: ISA Brown and ISA White layers under experimental conditions. 2. A total of 72 chickens from each of these breeds were randomly distributed to 4 pens to provide equal numbers of two replicate pens maintained as infected and control (uninfected). All chickens in infected groups were inoculated orally with 1 × 108 CFU (1 ml dose) of a field isolate of S. gallinarum, at the age of 8 and 16 weeks given over 5 consecutive days. Growth performance, clinical signs, gross pathological lesions and antibody responses were measured. 3. A significantly higher mortality was observed in the brown layers compared with the white layers, and clinical signs and mortality were absent in village chickens. However, a large number of birds with gross lesions and high antibody titres were detected in village chickens, indicating that birds had the disease subclinically. Commercial breeds had a significantly higher body weight, feed intake and feed conversion efficiency. 4. There was a significantly lower proportion of positive reactors in village chickens in the whole-blood agglutination test (35%) compared to brown (100%) and white (90%) layers even after the second inoculation. Uninfected birds were negative in all groups. The indirect enzyme-linked immunosorbent assay confirmed these observations. 5. These results suggest that the indigenous breed had superior natural resistance to S. gallinarum than the commercial breeds.

Effects of feeding the herb Borreria latifolia on the meat quality of village chickens in Malaysia.

An experiment was carried out to estimate the meat quality characteristics of village chickens (Gallus gallus) fed diets supplemented with dry leaves of Borreria latifolia (BL) used as a potential antioxidant source in chicken feed. In this study, 252 sexed 9-week-old village chickens with mean live body weight of 1,525.4 g for males and 1,254.1 g for females were divided into 7 groups (each group 18 birds) for each sex represented in 2 experiments. The first experiment was to evaluate the antioxidant activity of BL and the effect on meat quality through a comparison with Rosmarinus officinalis (RO); hence, 3 groups were conducted and included: T1 (control), basal diet without supplementation; T2, basal diet with 1% of BL; T3, basal diet with 1% of RO. T2 and T3 significantly affect pH value, lipid oxidation, cooking loss, and overall acceptability compared to T1, while no significant difference was observed between the dietary groups in respect of drip loss, color, tenderness, fatty acid profile, and meat composition. Furthermore, a significant effect of sex on lipid oxidation, pH, yellowness, and fatty acid profile was observed. There was no significant effect of sex on WHC, tenderness, lightness, redness, and sensory evaluation. A significant influence of postmortem aging period was detected on lipid oxidation, pH, tenderness, cooking loss, and redness. The obtained result in this study revealed a significance in the interaction of herb by sex in pH parameter and between herb and sex, herb by aging period, sex by aging period, and the herb by sex by aging period interactions with regard to lipid oxidation test. The second experiment was to estimate the effect of 3 different levels of BL on meat quality. Four groups were provided and involved: T1 (control), basal diet without supplementation; T2, basal diet with 1.5% of BL; T3, basal diet with 2% of BL; and T4, basal diet with 2.5% of BL. The result of this study showed a significant effect (P < 0.05) of the dietary groups on redness value, sensory evaluation, and lipid oxidation. A significant effect of sex on fatty acid profile and lipid oxidation was detected in addition a significant effect of postmortem aging period on redness, pH, tenderness, cooking loss, and lipid oxidation was noticed. There was no significance in the interaction between herb and sex through pH test while a significance in the interaction between herb and sex, herb  × aging period, sex × aging period, and herb × sex × aging period interactions with regard to lipid oxidation was found. It can be concluded that dietary supplementation of BL improved oxidative stability and enhanced meat quality.

Infection-interactions in Ethiopian village chickens.

Chickens raised under village production systems are exposed to a wide variety of pathogens, and current or previous infections may affect their susceptibility to further infections with another parasite, and/or can alter the manifestation of each infection. It is possible that co-infections may be as important as environmental risk factors. However, in cross-sectional studies, where the timing of infection is unknown, apparent associations between infections may be observed due to parasites sharing common risk factors. This study measured antibody titres to 3 viral (Newcastle disease, Marek's disease and infectious bursal disease) and 2 bacterial (Pasteurella multocida and Salmonella) diseases, and the infection prevalence of 3 families of endo- and ecto-parasites (Ascaridida, Eimeria and lice) in 1056 village chickens from two geographically distinct populations in Ethiopia. Samples were collected during 4 cross-sectional surveys, each approximately 6 months apart. Constrained ordination, a technique for analysis of ecological community data, was used to explore this complex dataset and enabled potential relationships to be uncovered and tested despite the different measurements used for the different parasites. It was found that only a small proportion of variation in the data could be explained by the risk factors measured. Very few birds (9/1280) were found to be seropositive to Newcastle disease. Positive relationships were identified between Pasteurella and Salmonella titres; and between Marek's disease and parasitic infections, and these two groups of diseases were correlated with females and males, respectively. This may suggest differences in the way that the immune systems of male and female chickens interact with these parasites. In conclusion, we find that a number of infectious pathogens and their interactions are likely to impact village chicken health and production. Control of these infections is likely to be of importance in future development planning.

Interdisciplinary communication of infectious disease research - translating complex epidemiological findings into understandable messages for village chicken farmers in Myanmar.

Improvement in animal disease control and prevention is dependent on several factors including farmers' uptake of new technologies and skills, particularly in developing countries. Extension is the means by which information about these technologies and skills is delivered to farmers, in order that they can use this knowledge to improve farming practices and their quality of life. This implies a shift from traditional methods to new science-based methods of production. However, in many developing countries farmers are illiterate and unable to understand written outcomes of scientific research. This paper summarizes approaches to communicate epidemiological findings and reports on experiences obtained from a research project in Myanmar, where results from epidemiological field investigations and intervention studies were 'translated' in an understandable manner to village communities. Rural chicken farmers were the central focus of this extension work and simple and sustainable methods to improve the health and production of scavenging chicken flocks were promoted. Unique extension materials transformed scientific outputs published in international journals into clear pictographic messages comprehendible by villagers, while maintaining country-specific, traditional, religious and public perspectives. Benefits, difficulties and pitfalls in using extension methods to communicate advice on preventive veterinary medicine measures in different cross-cultural settings are discussed and guidelines on how to distribute epidemiological research results to illiterate farmers are provided.