Losses In Poultry Research Articles

Kicking Newcastle Disease in Poultry: The Therapeutic Potential of a Herbal Blend of Christmas Melon, Aloe Vera, Chili, and Ash Extract

Newcastle Disease (ND) is a highly contagious viral infection that causes substantial mortality and economic losses in poultry, particularly in resource-limited settings like rural Uganda. This study explores the therapeutic potential of a herbal blend composed of Christmas melon, aloe vera, chili, and ash extract as an alternative or adjunct treatment for ND in poultry. Grounded in Integrative Medicine Theory (IMT), which emphasizes the synergy between conventional and traditional medical approaches, the study investigates the effectiveness of this herbal remedy in both treating and preventing ND. The results, based on personal case studies and a larger preventive trial, demonstrate a high recovery rate in affected birds and successful prevention in exposed flocks. Notably, six out of seven birds treated for ND showed full recovery, and no new ND cases emerged in the preventive trial involving 1,654 layers. However, a slight decrease in egg production was observed in treated layers, signaling a potential side effect that warrants further research. This study underscores the viability of herbal remedies in managing ND in areas where conventional treatments may be limited or inaccessible, providing a holistic, farmer-centered, and cost-effective solution to poultry health management in rural African settings. While promising, further research is needed to optimize dosage, assess long-term safety, and minimize adverse effects.

Treatment of avian pathogenic Escherichia coli infected broilers with aqueous extracts of Vernonia amygdalina in a challenge experiment

Colibacillosis, which is the leading cause of infectious diseases resulting in mortality and economic losses in poultry is triggered by Avian pathogenic Escherichia coli (APEC). Conventionally, antibiotic growth promoters (AGP) are incorporated into poultry diets to control infectious diseases and boost performance. However, because of the problem of antimicrobial resistance, synthetic antibiotics are now restricted or banned in animal production in some countries. In this study, we carried out a challenge experiment by inoculating healthy broilers with APEC (106 cfu/mL) by oral gavage on day 21 of the experiment and orally administering various treatments five days post challenge for five days; synthetic antibiotics (T2), Vernonia amygdalina (bitter leaf) extracts (T3), no treatment (T4), which were compared with the control that wasn’t challenged (T1). The experiment lasted for 42 days. Performance, hematological and histological studies were carried out. The results show that the challenged birds became diseased with the development of visible lesions, which ameliorated over time to varying extents with the use of synthetic antibiotics and Vernonia treatments. Overall, the performance of the birds with the use of Vernonia (T3) was comparable with that of synthetic antibiotics (T2) without eliciting any adverse hematological effects on the broilers. We therefore conclude that Vernonia can be safely used as a supplement for disease prevention in broiler chicken production

Detection and genetic characterization of chicken astrovirus and avian nephritis virus from hatchery to farm

ABSTRACT Astroviruses are an emerging hazard in hatchery management, and they are responsible for great economic losses in poultry farms worldwide. Chicken astrovirus (CAstV) and avian nephritis virus (ANV) are involved in decreased hatching rate, mainly due to embryo deaths, and pale and runted chicks too weak to hatch. Hatchery diseases occur through direct egg infection or vertical transmission. However, the specific role of astroviruses in these diseases is still largely unknown. To elucidate this a high-quality Piedmont hatchery (Italy) was selected based on farm clinical signs and the astrovirus serological positivity in the breeder farm. The viral dynamics were followed from the hatchery to the farm. The presence of both CAstV and ANV was monitored by evaluating the viral load in environmental and bird samples. Interestingly, both viruses were detected in chick yolk sacs at 18 days of incubation, and in the gut contents of 1-day chicks, whereas in the hatchery environmental samples, the eggs' external surfaces and the embryos’ intestinal contents were negative. The CAstV copy number was higher in bird tissues from the hatchery, while a greater ANV load was detected in bird tissues from the farm. Moreover, our data suggest vertical transmission of both viruses. Frequent co-infection was also observed in samples from both the hatchery and the farm. Lastly, the first Italian full-length genome and molecular characterization of ANV was obtained. RESEARCH HIGHLIGHTS Identified the role of the hatchery in astrovirus transmission. Sequenced the avian nephritis virus complete genome. Investigated tissue distribution of astrovirus from egg to chicks. Demonstrated co-infection of ANV/CAstV.

Prevalence and molecular characterization of circoviruses, astroviruses and hepadnaviruses in the avifauna of Kopački rit Nature Park, Croatia

Wild birds harbour a plethora of viruses but the research into them has mainly focused on those possessing zoonotic potential or causing significant economic losses in poultry, while others, such as astroviruses (AstVs), circoviruses (CV) and hepadnaviruses (HBV), remain neglected. In this study, 35 dead wild birds belonging to at least 21 species from five orders, collected in Kopački rit Nature Park were tested for the presence of AstVs, CV and HBV using PCR with subsequent nucleotide sequencing. A positive result was found in 18 birds. AstVs were found in three passerine birds, CV was detected in 11 birds belonging to Accipitriformes, Anseriformes, Charadriiformes, Passeriformes or Piciformes orders while HBV was found in four birds belonging to either Anseriformes or Passeriformes. More than one virus was found in a single species (AstV and CV) although in two separate birds. Phylogenetic analysis revealed divergent AstVs somewhat similar to different unclassified AstVs detected in undetermined birds. One AstV shared the highest homology with a bat AstV (98.77%). All CV found in the study resemble avian CV, but none of them clusters in the phylogenetic tree with any avian or avian-like CV sequence. All four HBV sequences detected in this study cluster within the duck HBV group, indicating unusual interspecies transmission to cohabitating passerines. The results of this study represent an important contribution to knowledge about the prevalence, ecology and epidemiology of AstVs, CV and HBV in different wild bird species, indicating the need for further studies, including whole genome sequencing of the detected viruses.

Genome Sequences and Characterization of Chicken Astrovirus and Avian Nephritis Virus from Tanzanian Live Bird Markets.

The enteric chicken astrovirus (CAstV) and avian nephritis virus (ANV) are the type species of the genus Avastrovirus (AAstV; Astroviridae family), capable of causing considerable production losses in poultry. Using next-generation sequencing of a cloacal swab from a backyard chicken in Tanzania, we assembled genome sequences of ANV and CAstV (6918 nt and 7318 nt in length, respectively, excluding poly(A) tails, which have a typical AAstV genome architecture (5'-UTR-ORF1a-ORF1b-ORF2-'3-UTR). They are most similar to strains ck/ANV/BR/RS/6R/15 (82.72%) and ck/CAstV/PL/G059/14 (82.23%), respectively. Phylogenetic and sequence analyses of the genomes and the three open reading frames (ORFs) grouped the Tanzanian ANV and CAstV strains with Eurasian ANV-5 and CAstV-Aii viruses, respectively. Compared to other AAstVs, the Tanzanian strains have numerous amino acid variations (substitutions, insertions and deletions) in the spike region of the capsid protein. Furthermore, CAstV-A has a 4018 nt recombinant fragment in the ORF1a/1b genomic region, predicted to be from Eurasian CAstV-Bi and Bvi parental strains. These data should inform future epidemiological studies and options for AAstV diagnostics and vaccines.

SteE enhances the colonization of Salmonella Pullorum in chickens

Salmonella pullorum (S. pullorum) is the causative agent of pullorum disease and results in severe economic losses in poultry, and can long-term survival by colonizing host organs. steE is an effector protein secreted by Salmonella pathogenicity island 2. It is not clear in vivo for the colonization of Salmonella. To investigate the role of steE on the colonization of S. Pullorum in the principal organs of chicken, we used S. pullorum and S. pullorum ΔsteE strains immunized chickens, respectively. The results of the virulence assay showed that the LD50 of S. pullorum ΔsteE was 22.8 times higher than that of S. pullorum in chickens. The colonization experiment of bacteria showed that the overall change trend of the number of S. pullorum and S. pullorum ΔsteE strains were similar in chicken liver, spleen, heart, bursa, and cecum, which increased first and then decreased. However, the deletion of steE caused significantly reduced colonization, pathological change, and virulence of S. pullorum in a chicken infection model. Our findings provide exciting insights into the pathogenic mechanism and live attenuated vaccine associated with steE in S. pullorum.

Molecular detection of highly pathogenic avian influenza H5N8 in commercial broiler chicken farms from 2019 to 2022

Highly pathogenic avian influenza (HPAI) is a serious viral infection that causes massive economic losses in poultry. The current study investigated the HPAI virus prevalence in commercial broiler chicken flocks from 2019 to 2022. Organ samples, including trachea, cecal tonsils, spleen, brain, as well as tracheal and cloacal swabs, were harvested from 111 problematic broiler chicken flocks that suffered from variable mortalities accompanied with respiratory signs (103 H5-vaccinated and 8 nonvaccinated flocks) in Egypt during the observation duration. Molecular tools were used to analyze the samples, including real-time reverse transcription-polymerase chain reaction (rRT-PCR) and sequence analysis of some PCR positive strains. The results indicated that 24 flocks were positive for HPAI H5N8, representing 21.6%, with 22.3% (23/103) prevalence and 12.5% (1/8) detection in vaccinated and nonvaccinated flocks, respectively, and they were almost detected in the autumn and winter seasons. Phylogenetic evaluation of the hemagglutinin (HA) gene showed that the 6 Egyptian strains were clustered in clade 2.3.4.4b and allocated into 2 groups (I and II). The samples recovered in 2019 were clustered in new subgroup A, and samples recovered in 2020 to 2022 were clustered in new subgroup B with 10 nucleotide mutations (R72S, A83D, T140A). In conclusion, HPAI H5N8 is a serious threat even in vaccinated birds; to control such problems, periodic molecular monitoring with vaccine efficacy evaluation and the use of preventive strategies are recommended.

Evaluation of inactivated vaccine's Antibody response to different H9N2 Vaccination programs with Hemagglutination Inhibition (HI) assay

Influenza is a significant poultry disease that can also affect humans, making it a zoonosis. The World Health Organization (WHO) and Contagious Diseases Organization (CDO) have increasingly focused on this disease in recent years. The isolation of influenza, particularly H5N1 and H9N2 subtypes, has dramatically increased, posing a global epidemic threat and causing deaths in various communities, as well as substantial losses in poultry. In response, using inactivated avian influenza vaccines has become common in controlling the disease. A comprehensive study was conducted in broiler farms in Iran to address the importance of influenza control through vaccination. The study involved 50,000 broiler chicks divided into seven groups with different vaccination programs. The groups were vaccinated at different ages and received varying vaccine doses. Serum samples were collected weekly and analyzed for antibody titers using hemagglutination inhibition (HI). The results showed that the groups vaccinated after seven days of age had a more consistent immune response and higher antibody titers than those vaccinated at a younger age or with only one dose. Early vaccination before seven days of age did not effectively stimulate the desired antibody response or achieve expected titers for the H9N2 influenza virus. Therefore, vaccinating older chicks improved immunity and flock protection more effectively. This study highlights the importance of vaccination strategies in controlling influenza in broiler farms. It provides valuable insights into the optimal timing and dosage of vaccines to enhance the immune response and protect against the H9N2 subtype. Implementing appropriate vaccination programs can contribute to mitigating the impact of influenza on poultry and reducing the risk of transmission to humans, ultimately safeguarding both animal and public health.

Prevalence of Salmonella in poultry slaughterhouses located in Tripoli, Libya.

Salmonella is a leading cause of severe economic losses in poultry and foodborne illness in humans worldwide. The aim of this study was to determine the prevalence and multidrug resistance of Salmonella Enteritidis (S. Enteritidis) in several chicken abattoirs in Tripoli, Libya. The study includes the South, East, and West regions of Tripoli. Each region was assigned five slaughterhouses. Each chicken slaughterhouse was visited three times to collect samples. Five samples were taken at random from the neck skin, crop, and spleen. The total number of samples collected from all regions was 675. Bacterial isolation and identification, as well as antibiotic sensitivity testing, were performed on these samples. Salmonella spp. was found to be 15% prevalent, and S. Enteritidis was found to be 7% prevalent. The south region of Tripoli had the highest S. Enteritidis (9%), while the west region had the highest Salmonella spp. (22%). Salmonella prevalence increased significantly (p < 0.01) higher in the spleen (13%) as compared with the crop (5%) and neck (7%). Based on bacterial resistance pattern, Salmonella spp. isolated from the spleen had the highest multiple antibiotic resistance (MAR) index of 0.86 in the south region followed by MAR indexes of 0.8 and 0.46 in the West and East, respectively. Isolation of Salmonella from the spleen may indicate chickens' systemic infection and failure to control the most important microbe for public health. Thus, the control measures have to be revised and a national Salmonella control program should be put in place urgently.

Genetic Analyses of Response of Local Ghanaian Tanzanian Chicken Ecotypes to a Natural Challenge with Velogenic Newcastle Disease Virus.

Newcastle disease is a devastating poultry disease that often causes significant economic losses in poultry in the developing countries of Africa, Asia, as well as South and Central America. Velogenic Newcastle disease virus (NDV) outbreaks are associated with high mortalities, which can threaten household livelihoods, especially in the rural areas, and lead to loss of high-quality proteins in the form of meat and eggs, as well as household purchasing power. In this study, we exposed unvaccinated Ghanaian and Tanzanian chickens of six local ecotypes to velogenic NDV strains, measured NDV response traits, sequenced their DNA on a genotyping-by-sequencing platform, and performed variance component analyses. The collected phenotypes included: growth rates (pre- and post-exposure); lesion scores (gross lesion severity) in the trachea, proventriculus, intestine, and cecal tonsils; natural antibody levels; anti-NDV antibody levels at 7 days post exposure (dpe); tear and cloacal viral load at 2, 4, and 6 dpe; and survival time. Heritability estimates were low to moderate, ranging from 0.11 for average lesion scores to 0.36 for pre-exposure growth rate. Heritability estimates for survival time were 0.23 and 0.27 for the Tanzanian and Ghanaian ecotypes, respectively. Similar heritability estimates were observed when data were analyzed either separately or combined for the two countries. Survival time was genetically negatively correlated with lesion scores and with viral load. Results suggested that response to mesogenic or velogenic NDV of these local chicken ecotypes could be improved by selective breeding. Chickens that are more resilient to velogenic NDV can improve household livelihoods in developing countries.

Preventive effect of on-farm biosecurity practices against highly pathogenic avian influenza (HPAI) H5N6 infection on commercial layer farms in the Republic of Korea during the 2016-17 epidemic: A case-control study

Highly pathogenic avian influenza virus (HPAIv) H5N6 has destructive consequences on the global poultry production system. Recently, a growing number of layer farms have been heavily damaged from the HPAIv epidemic due to the increased virulence of the virus and the intensification of the production system. Therefore, stakeholders should implement effective preventive practices at the farm level that are aligned with contingency measures at the national level to minimize poultry losses. However, numerous biosecurity protocols for layer farm workers to follow have been developed, impeding efficient prevention and control. Furthermore, the effectiveness of biosecurity practices varies with the geographical condition and inter-farm contact structures. Hence, the objective of our study was to examine the preventive effect of five biosecurity actions commonly practiced at layer farms in the Republic of Korea against HPAIv H5N6: (i) fence installation around a farm, ii) rodent control inside a farm; iii) disinfection booth for visitors for disinfection protocols, iv) an anterior room in the sheds before entering the bird area and v) boots changes when moving between sheds in the same farm. We conducted a case-control study on 114 layer case farms and 129 layer control farms during the 2016-17 HPAI epidemic. The odds ratios for five on-farm biosecurity practices implemented in those study groups were estimated as a preventive effect on the HPAI infection with covariates, including seven geographical conditions and three network metrics using Bayesian hierarchical logistic regression and geographical location weighted logistic regression. The results showed that the use of a disinfection booth for personnel reduced the odds of HPAIv H5N6 infection (adjusted odds ratio [AOR] = 0.002, 95 % credible interval [CrI] = 0.00007 - 0.025) with relatively small spatial variation (minimum AOR – maximum AOR: 0.084−0.263). Changing boots between sheds on the same farm reduced the odds of HPAIv H5N6 infection (AOR = 0.160, 95 % CrI = 0.024−0.852) with relatively wide spatial variation (minimum AOR – maximum AOR = 0.270−0.688). Therefore, enhanced personnel biosecurity protocols at the farm of entry for layer farms is recommended to effectively prevent and respond to HPAIv H5N6 infection under different local condition. Our study provides an important message for layer farmers to effectively implement on-farm biosecurity actions against HPAIv H5N6 infection at their farms by setting priorities based on their spatial condition and network position.

Biochemical and molecular mechanisms of acaricide resistance in Dermanyssus gallinae populations from Turkey

The poultry red mite, Dermanyssus gallinae, is the most important blood sucking ectoparasite of egg laying hens and causes economic losses in poultry farms worldwide. Although various management methods exist, the control of poultry red mites (PRMs) mainly relies on acaricides such as pyrethroids and organophosphates (OPs) in many regions of the world. However, repeated use of these synthetic chemicals has resulted in resistance development causing control failure of PRM. In this study, we investigated acaricide resistance mechanisms of Turkish PRM populations. First, we obtained the COI sequence from 30 PRM populations from different regions in Turkey and identified four different COI haplotypes. Toxicity assays showed that four field-collected PRM populations were highly resistant to the pyrethroid alpha-cypermethrin, with resistance ratios (RRs) varying between 100- and 400-fold, while two of these populations had a RR of more than 24-fold against the OP acaricide phoxim. Biochemical assays showed a relatively higher activity of glutathione-S-transferases and carboxyl-cholinesterases, two well-known classes of detoxification enzymes, in one of these resistant populations. In addition, we also screened for mutations in the gene encoding the voltage-gated sodium channel (vgsc) and acetylcholinesterase 1 (ace-1), the target-site of pyrethroids and OPs, respectively. In all but two PRM populations, at least one vgsc mutation was detected. A total of four target-site mutations, previously associated with pyrethroid resistance, M918T, T929I, F1534L, F1538L were found in domain II and III of the VGSC. The T929I mutation was present in the vgsc of almost all PRM populations, while the other mutations were only found at low frequency. The G119S/A mutation in ace-1, previously associated with OP resistance, was found in PRM for the first time and present in fourteen populations. Last, both alive and dead PRMs were genotyped after pesticide exposure and supported the possible role of target-site mutations, T929I and G119S, in alpha-cypermethrin and phoxim resistance, respectively. To conclude, our study provides a current overview of resistance levels and resistance mutations in Turkish PRM populations and might aid in the design of an effective resistance management program of PRM in Turkey.

Avian Mycoplasma gallisepticum and Mycoplasma synoviae: Advances in diagnosis and control

Both of Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) infections are the most common Mycoplasma infection in domestic poultry. The disease is associated with economic losses in poultry. MG and MS are commonly spread within chickens (Gallus gallus domesticus) and turkeys (Meleagris gallopavo domesticus) flocks; however, they are frequently isolated from quails (Coturnix coturnix) and several avian species. Diagnosis of MG or MS infections is confirmed by isolating the organism in a cell-free medium or directly detecting its DNA in infected tissues or swab samples. Serological tests are also widely used for diagnosis. However, advances in molecular biology represented a rapid and sensitive alternative to the traditional culture methods requiring specialized techniques and sophisticated reagents. Several Mycoplasma molecular diagnostic tests are implemented: including polymerase chain reaction (PCR), Random Amplified Polymorphic DNA (RAPD), arbitrary primed polymerase chain reactions (AP‐PCR), and Multiplex real-time polymerase chain reaction (Multiplex MGMS). Current control practices against Mycoplasma infection include intense biosecurity, biosurveillance, medication, and vaccination. However, the egg-borne nature of avian Mycoplasma infection complicates controlling the infection. This review focuses on the advances in diagnosis and control of avian Mycoplasma infection, especially MG and MS infections.

Analysis of the prevalence of selected qualitative deviations in slaughter poultry in Poland in 2016-2019

The aim of the study was to analyse the results of official post-mortem inspections of hens, chickens, turkeys, ducks and geese carried out in poultry slaughterhouses in Poland by the Veterinary Inspection in 2016–2019. The data from the sanitary and veterinary survey were taken from annual reports drawn up by the General Veterinary Inspectorate (RRW-6) in 2016–2019. During this period, nearly 900 million hens, 206 million turkeys, 122 million ducks, 63 million geese, and over 5.7 billion slaughter chickens were slaughtered under veterinary supervision. Symptoms or lesions were found in almost 2.6 million hens (0.29%), 1.8 million turkeys (0.86%), 792 thousand ducks (0.65%), 185 thousand geese (0.29%) and over 27.3 million chickens (0.48%). The main causes of confiscation were emaciation (0.14%), incomplete loss of blood (0.06%), and septicaemia and pyaemia (0.07%). Quality deviations in the form of emaciation or incomplete loss of blood of slaughtered animals indicate shortcomings during rearing or transport of animals for slaughter. Therefore, there may be legitimate concerns about the appropriate level of animal welfare. The size of overall losses in poultry is determined by the disqualification of chickens, due to slaughter size in comparison with other species. The confiscation of such a large number of chickens deserves greater attention to determine the causes involved.

The effect of different levels of ozone and ozonized water on biogenic amines in broiler chicken meat by HPLC

Biogenic amines (BA) are low molecular weight substances, formed mainly by decarboxylation of specific amino acids present in food through the action of enzymes during storage produced by some microorganisms, this fact can be used to relate BA as a bacteriological quality indicator. This study was aimed to evaluate the effects of an experimental ozone gaseous treatment and production of the biogenic amines’ putrescine and cadaverine. Amines were extracted with perchloric acid, derivative with dansyl chloride, separated using a reversed-phase high-performance liquid chromatographic method, and detected by fluorescence. The results showed that during the 7 days, the amount of putrescine had increased in all four experimental treatments. The highest increase was related to Group 1. The Duncan post hoc test showed that the highest amount of Cadaverine after killing was related to Group 1 (control) with 88.85 mg/kg and the lowest value for Group 2 is 12.03 mg/kg. Also, the results of Duncan's post hoc test for comparing Cadaverine seven days after slaughter among experimental treatments showed that the highest amount of Cadaverine after slaughter was related to Group 1 (control) with 135.6 mg/kg and the lowest value for the group is 94.83 mg/kg. The results of the test to compare the pH of the treatments showed that the highest amount of pH is for Group 4 and the lowest value for Group 1 (control). The results of this study showed that with increasing ozone gas concentration and decreasing the concentration of water, the biogenic amines’ putrescine and cadaverine in frozen poultry meat decreased. Putrescine and cadaverine levels appeared to be useful to control the effectiveness of the ozone treatment on meat quality and may be useful as a quality index to highlight the loss of poultry meat freshness.

Effect of infectious bronchitis and Newcastle disease vaccines on experimental avian influenza infection (H9N2) in broiler chickens

Despite the fact that H9N2 avian influenza virus (AIV) is considered a low-pathogenic agent, frequent outbreaks of this subtype have caused high mortality and economic losses in poultry farms around the world including Iran. Coinfection with a respiratory pathogen or environmental factors may explain the exacerbation of H9N2 AIV infection. In this study, the role of infectious bronchitis (IB) vaccines (H120 and 4/91) and Newcastle disease (ND) vaccines (B1 and LaSota) on experimental H9N2 AIV infection was investigated in 180 broiler chickens allotted into 6 groups (n=30). At the age of 18 days, groups 3 and 4 received H120 and 4/91 infectious bronchitis live vaccines (IBLVs) and groups 5 and 6 received B1 and LaSota Newcastle disease live vaccines (NDLVs), respectively. At the age of 20 days, all birds in the experimental groups except the negative control group (group 1), were inoculated intra-nasally with H9N2 AIV. After the inoculation, clinical signs, gross and microscopic lesions, and viral detection were examined. The results of this study revealed that clinical signs, gross and microscopic lesions were more severe in the AIV challenged groups which had been previously vaccinated with IB vaccines. In addition, AI viral RNA from tracheal and faecal samples in IB vaccinated birds were recovered at a higher rate. Moreover, in the 4/91 IB vaccinated group, the AI virus shedding period was longer than the other challenged groups. In conclusion, infectious bronchitis live vaccines (IBLVs) exacerbated the H9N2 AIV infection; also, 4/91 IBLV extended AI virus shedding period and increased the recovery rate of AI virus from feaces. However, the coinfection of Newcastle disease live vaccines (NDLVs) had no considerable adverse effects on AIV infection in broiler chickens.

Meat Value Chain Losses in Iran.

To stop hunger, reducing food losses is a potential movement towards saving food. A large portion of these losses could be avoided and reduced through the improved food chain in many countries. Raising awareness on how and where food losses occur will help recovering foods such as meat by identifying solutions and convincing people to implement those solutions. This, in turn, will lead to private and public efforts to recover meat that might be otherwise wasted. After highlighting the importance of food saving benefits and relevant statistics, this paper explains the possible ways to reduce meat loss and waste in abattoirs and presents a framework for prevention according to the estimates of meat losses in Iran meat supply. The current article answers the questions of where do we have the meat loss in Iran and what approaches are most successful in reducing losses in the meat industry. The national average loss and waste in meat production are about 300,000 metric tonnes (about 15%). Many segments and players are involved with this huge amount of losses in the meat value chain, a large portion of these losses could be avoided and reduced by about 25% through using by-products with the mechanization of design and manufacturing. The production amount of mechanically deboned meat (MDM) is 105,091,000 kg, concluding the major waste (88.33%) of total poultry losses. Ensuring appropriate actions by exploiting the full potential of engaged Iranian associations and institutes is considered to reduce the losses.

Immunization of turkeys with a DNA vaccine expressing the haemagglutinin gene of low pathogenic avian influenza virus subtype H9N2

Low pathogenic avian influenza H9N2 is still circulating in the Middle East causing respiratory manifestations and severe economic losses in poultry. In the present study, an H9 plasmid-based DNA vaccine targeting the HA gene of H9N2 A/CK/Egypt/SCU8/2014 was developed and evaluated in turkeys. The full length of HA was cloned into vector plasmids under the control of a cytomegalovirus promoter. The in-vitro expression of the recombinant HA was demonstrated in HeLa cells transfected with the plasmids pVAX1-H9 or pCR-H9 using western blot and Immunofluorescent assay (IFA). The efficacy of pVAX-H9 and pCR- H9, naked or saponin-adjuvanted, was evaluated in turkey poults at 3 weeks and challenged with A/CK/Egypt/SCU8/2014 (106 EID50/bird at 3 weeks post-vaccination. The efficacy was assesses based on virus shedding, oropharyngeal and cloacal, as well as seroconversion using haemagglutination inhibition (HI) test. All immunized birds showed high HI antibody titers (7–8 log2) at 3 weeks post-vaccination. None of the birds vaccinated with naked or saponin-adjuvanted pVAX-H9 or pCR-H9 showed any clinical signs. The pVAX-H9 and pCR-H9 alone did not prevent cloacal and oropharyngeal virus shedding, however, saponin-adjuvanted pVAX1-H9 and pCR-H9 prevented cloacal and oropharyngeal virus shedding at 3 and 5 days post challenge, respectively. In conclusion, DNA vaccination with pVAX1-H9 and pCR-H9 could protect turkey from the H9N2 virus, but vaccination regimes need to be improved.

Expression of blaCTX-M2 and invA genes of Salmonella Heidelberg isolated from poultry by Qpcr

Salmonellosis is a disease caused by a bacterium Salmonella, a gram negative bacilli found in many environments, responsible for significant economic losses in poultry, and of great impact on public health. Among more than 2500 serovars, S. Heidelberg seems to be more invasive causing disease of greater severity than other serovars. The objective of this study was to investigate, through real-time PCR (qPCR), differences in the expression of a virulent gene (invA) and an antibiotic resistance gene (blaCTXM-2) of S. Heidelberg isolated from poultry meat (slaughterhouses) and drag swabs (field). Even though all isolates showed the presence of the invA gene, there were differences in the expression among the isolates, where isolates from the field showed greater expression of invA compared to samples isolated from meat products. On the other hand, isolates from the slaughterhouses showed greater expression of the blaCTX-M2 than those isolated from field samples.

Evaluating two approaches for using positive control in standardizing the avian influenza H5 reverse transcription recombinase polymerase amplification assay

Highly pathogenic avian influenza H5N1 virus causes heavy losses in poultry farms worldwide. Molecular diagnostic techniques like RT-PCR and real-time RT-PCR are considered the gold standard for identification of H5 influenza viruses in clinical samples. These techniques are hampered by the need of well-equipped laboratories, large space requirement, and relatively long time-to-result. Recombinase polymerase amplification (RPA) assay represents an excellent alternative to PCR since it is more simple, rapid, economic, and portable. Reverse transcription RPA (RT-RPA) assay was recently developed for sensitive and specific detection of H5N1 virus in 6–10 min. To ensure the accuracy of the developed assay, two approaches for using a positive control were evaluated in this study. These approaches included: 1) all-in-one (internal positive control; IPC), 2) two-tubes-per-one-sample (external positive control; EPC). Sigma virus (SIGV) RNA and turkey mitochondrial DNA were tested as positive controls in both approaches. For all-in-one approach, both targets (H5 and IPC) were strongly inhibited. In contrast, very good amplification signals were obtained for the two types of EPC with no effect on the analytical sensitivity and specificity of H5 RT-RPA assay in two-tubes-per-one-sample approach. The performance of EPC-based H5 RT-RPA was further validated using 13 tracheal swabs. The results were compared to real-time RT-PCR and proved superior specificity in detecting H5N1 but not H5N8 viruses. Inclusion of EPC did not affect the aptitude of both assays in terms of sensitivity, specificity and reproducibility. In conclusion, the two-tubes-per-one-sample approach was more reliable to control the false negative results in H5 RT-RPA assay.