Pharmaceutical activity of sappan wood extract (Caesalpinia sappan L.) for treating Escherichia coli infection in piglets.

Background: Escherichia coli infection is one of the major diarrheal diseases resulting in the loss of pigs at a young age. Aim: This research investigated the antimicrobial activity of Caesalpinia sappan wood extract against E. coli infection as an antibiotic replacement. Methods: E. coli was cultured from diarrheal piglets and then used to find the minimal inhibition concentration (MIC). Caesalpinia sappan wood extract (500 mg/kg) was used for the treatment of diarrheal piglets compared to antibiotics (enrofloxacin 5 mg/kg) by oral administration. Another three groups of diarrheal piglets were used supplemented feed with 1% and 2% extract compared with commercial feed. Subsequently, E. coli enumeration, fecal shape, fecal color, and growth rate were recorded from day 1 to 7. Results: Based on the results, C. sappan wood extract could inhibit E. coli growth at a MIC of 16-34 mg/ml. The number of colonies did not significantly differ between C. sappan wood extract and enrofloxacin treatment groups. A supplemented feed with 1% and 2% C. sappan wood extract could improve the fecal shape and fecal score compared to the control group, albeit only in suckling pigs. There were significant differences from the control group on days 4, 5, 6, and 7 (p < 0.05). However, the average daily gain did not significantly differ among the three groups. Conclusion: The results indicate that C. sappan wood extract could improve diarrheal signs in suckling pigs and can be used as a replacement for antibiotics for organic pig production.

Phage cocktail administration to reduce Salmonella load in broilers.

Salmonella is a serious foodborne pathogen that can cause gastrointestinal disease through the consumption of contaminated foods; including poultry meat. Salmonella is commonly present in the intestinal tract of poultry and farm environments, posing a potential risk of contamination during the processing of poultry meat. This study was a continuation in evaluating the effects of our previously developed phage cocktail targeting Salmonella at large-scale trials in commercial broiler farms, in which this cocktail considerably lowered Salmonella colonization in the gut of broilers. The phage cocktail given to broilers showed resistance to temperatures of up to 65 °C (> 60% survivability), pH ranging from 2 to 12 (> 96% survivability), 0.5 to 15% (w/v) NaCl (> 98% survivability), chlorine up to 0.5% (v/v) (53% survivability), and chlorine neutralizer (100% survivability). In the animal challenge study, phage treatments, designed as "prevention" and "exclusion" programs, could control Salmonella on day 20 and 32 of the experiment, respectively; as indicated by the absence of Salmonella detection in cloacal swabs from broilers (0% prevalence). In the commercial-scale trial I, Salmonella was not detected in the phage-treated group from cloacal swabs, boot cover swabs, and bedding material samples after 16 days (0% prevalence) of phage administration. In the commercial-scale trial II, phage treatment extended the Salmonella control period in broilers during a 40-day growout period. In summary, a phage cocktail demonstrated high efficiency in controlling various serovars of Salmonella historically linked to contamination on these broiler farms. Phage cocktail application offers an effective, alternative to enhance food safety within the poultry value chain, protecting consumers and as well as the economic sustainability of the poultry sector.

Prevalence of Zoonotic Intestinal Parasites in Meat Goats in Southern Thailand.

Background: Intestinal parasites not only lead to poor goat health and productivity but can also affect human health. We investigated the distribution of zoonotic intestinal parasites among goats in southern Thailand to assess the human health risks. Materials and Methods: Overall, 561 fecal samples were collected randomly from goats in farms (55) in Songkhla, Satun, Pattalung, Pattani, Yala, and Narathiwat provinces between January 2020 to January 2022. Parasite eggs were detected via the standard protocol formalin ether concentration technique (FECT). Results: Samples containing one or more intestinal parasites accounted for 97.33% (543/561) of the total samples, and the percentage of zoonotic parasite was 80.04% (449/561). Among the positive samples, two types of zoonotic helminths were identified, Strongylid egg type (79.32%) and Fasciola spp. (2.14%). Strongylid egg type was highly abundant in samples from Satun (84.62%), Songkhla (82.74%), Pattalung (79.31%), and Yala (77.27%). Fasciola spp. was mostly found in samples form Pattalung (13.79%) and Songkhla (1.12%). Eimeria spp. was the mostly identified protozoa 25.67%. It was mostly identified in samples from Narathiwat (75%), Pattani (41.9%), and Songkhla (31.41%). Mixed infections accounted for 40.82% of the total with 33 patterns. It was found that Strongylid egg types occurred in 28 patterns (84.85%). Strongylid egg types-Eimeria spp.-pattern was the most commonly found. Conclusion: Based on the results, zoonotic helminth infection was found together with widespread and high diversity of intestinal parasites infection in meat goats, which affect animal health and may pose a risk to human health. Mass deworming programs with a proper anthelmintic drugs and good agricultural practices need to be encouraged by the government so that people who contact with goats should practice good hygiene behavior. Farm management should be applied by intensive goat raising with pasture circulating to reduce parasite egg contamination. Moreover, fresh fertilizers from goat faces should be avoided.

Tick-borne Pathogen Detection and Its Association with Alterations in Packed Cell Volume of Dairy Cattle in Thailand.

Tick-borne diseases (TBDs) massively impact bovine production. In endemic countries, animals are often subclinically infected, showing no signs of the illness. Anemia is a hallmark of TBDs, but there is inadequate information on its presence in infected Thai cattle. In the present study, 265 cattle from four provinces in Thailand were surveyed to identify tick-borne pathogens (TBPs) and to evaluate the changes in the packed cell volume (PCV) values associated with detection. Microscopy and polymerase chain reaction (PCR) were also compared for TBP detection. Babesia/Theileria/Hepatozoon was detected in 33.58% (89/265) of the cattle samples. Specifically, Babesia bovis (9/265), B. bigemina (12/265), Theileria orientalis (62/265), and Anaplasma marginale (50/265) were identified using species-specific assays. Significant decreases in the mean PCV levels were observed in cattle that were positive for at least one TBP (p < 0.001), Babesia/Theileria/Hepatozoon (p < 0.001), T. orientalis (p < 0.001), and A. marginale (p = 0.049). The results of PCR and microscopy for the detection of TBPs suggested slight and fair agreement between the two detection tools. The present findings contribute to a better understanding of TBDs in the field and shall facilitate the formulation of effective control for TBDs in Thailand.

Prevalence and Virulent Gene Profiles of Sorbitol Non-Fermenting Shiga Toxin-Producing Escherichia coli Isolated from Goats in Southern Thailand.

Shiga toxin-producing Escherichia coli (STEC) is the pathogenic E. coli causing disease in humans via the consumption or handling of animal food products. The high prevalence of these organisms in ruminants has been widely reported. Among STECs, O157 is one of the most lethal serotypes causing serious disease in humans. The present study investigated the prevalence of sorbitol non-fermenting STECs in goats reared in the lower region of southern Thailand and described the virulent factors carried by those isolates. Sorbitol non-fermenting (SNF)-STECs were found in 57 out of 646 goats (8.82%; 95% CI 6.75% to 11.28%). Molecular identification revealed that 0.77% of SNF-STEC isolates were the O157 serotype. Shiga toxin genes (stx1 and stx2) and other virulent genes (i.e., eaeA, ehxA, and saa) were detected by molecular techniques. The presence of stx1 (75.44%) was significantly higher than that of stx2 (22.81%), whereas 1.75% of the total isolates carried both stx1 and stx2. Most of the isolates carried ehxA for 75.44%, followed by saa (42.11%) and eaeA (12.28%). In addition, 21.05% of STEC isolates did not carry any eaeA, ehxA, or saa. The first investigation on SNF-STECs in goat was conducted in the lower region of southern Thailand. The present study revealed that goats could be one of the potential carriers of SNF-STECs in the observing area.

Oral Administration of a Phage Cocktail to Reduce Salmonella Colonization in Broiler Gastrointestinal Tract-A Pilot Study.

Salmonella contamination in poultry meat products can lead to serious foodborne illness and economic loss from product recalls. It is crucial to control Salmonella contamination in poultry from farm to fork. Bacteriophages (phages) are viruses of bacteria that offer several advantages, especially their specificity to target bacteria. In our study, three Salmonella phages (vB_SenS_KP001, vB_SenS_KP005, and vB_SenS_WP110) recovered from a broiler farm and wastewater treatment stations showed high lysis ability ranging from 85.7 to 96.4% on over 56 serovars of Salmonella derived from several sources, including livestock and a broiler farm environment. A three-phage cocktail reduced S. Enteritidis and S. Typhimurium, in vitro by 3.9 ± 0.0 and 3.9 ± 0.2 log units at a multiplicity of infection (MOI) of 103 and 3.8 ± 0.4 and 4.1 ± 0.2 log units at MOI of 104 after 6 h post-phage treatment. A developed phage cocktail did not cause phage resistance in Salmonella during phage treatments for three passages. Phages could survive under simulated chicken gastrointestinal conditions in the presence of gastric acid for 2 h (100.0 ± 0.0% survivability), bile salt for 1 h (98.1 ± 1.0% survivability), and intestinal fluid for 4 h (100 ± 0.0% survivability). Each phage was in the phage cocktail at a concentration of up to 9.0 log PFU/mL. These did not cause any cytotoxicity to human fibroblast cells or Caco-2 cells as indicated by the percent of cell viability, which remained nearly 100% as compared with the control during 72 h of co-culture. The phage cocktail was given to broilers raised in commercial conditions at a 9 log PFU/dose for five doses, while naturally occurring Salmonella cells colonized in the gastrointestinal tract of broilers were significantly reduced as suggested by a considerably lower Salmonella prevalence from over 70 to 0% prevalence after four days of phage treatment. Our findings suggest that a phage cocktail is an effective biocontrol agent to reduce Salmonella present in the guts of broilers, which can be applied to improve food safety in broiler production.

Isolation and in vitro cultivation of Trypanosoma evansi Thai strains.

Trypanosoma evansi is a flagellate protozoan parasite responsible for "surra". To generate T. evansi antigens for serodiagnosis, parasites are generally propagated in laboratory animals before isolation. The alternation of animal models using axenic cultivation systems to produce trypomastigotes of various Trypanosoma species is currently available but has never been applied in Thailand. The isolation protocol for separation of live T. evansi trypomastigotes from animal blood components before in vitro cultivation has not been clearly documented. This study focused on validation of trypomastigote isolation method, in vitro cultivation of T. evansi Thai strains, and its virulence ability in vivo. In this study, two strains of T. evansi collected from Thailand were used. Trypanosoma evansi trypomastigotes were propagated in mice, and three different isolation methods, including: low-speed centrifugation, high-speed centrifugation, and ion exchange chromatography using diethylaminoethyl (DEAE) cellulose (or DE52), were compared. Four solutions of in vitro cultivation media, two different in vitro cultivation containers, and different trypomastigote densities for initiation of in vitro culture were compared. Virulence test using in vitro-adapted parasite for 100 days was conducted in vivo. The results showed that the DE52 isolation method was suitable for separation of live T. evansi trypomastigotes from animal blood components before conducting in vitro cultivation. Trypanosoma evansi Thai strains were successfully cultivated and multiplied in HMI-9 Solution I using 25 cm2 flasks and 12-well plates. The parasite was growing slowly at the initiation of in vitro culture for 15-16 days, and then rapidly increased to 10, 20, 50, 100, and 200 folds, approximately. The doubling times were varied from 11.95 ± 8 h to 41.18 ± 4.29 h in vitro. The maximum densities have reached from 0.14 × 106 to 4.63 × 106 trypomastigotes/ml. Virulence test showed that the in vitro-cultivated T. evansi was virulent in mice. In conclusion, T. evansi Thai strains were successfully isolated and cultivated in vitro for the first time. The isolation and in vitro cultivation protocols were clearly provided. The benefit of using the in vitro cultivation system helps in the production of T. evansi antigen, and replacing the use of experimental animals. It is also useful for the development of diagnostic tests in the future.

Natural infection of leptospirosis and melioidosis in long-tailed macaques (Macaca fascicularis) in Thailand.

This study aimed to determine the incidence of leptospirosis and melioidosis in long-tailed macaques (Macaca fascicularis) in Thailand. Serum samples from 223 monkeys were subjected to the Lepto Latex Test and indirect hemagglutination (IHA) test to detect antibodies against Leptospira spp. and Burkholderia pseudomallei. The microagglutination test (MAT) was used to identify serovars of Leptospira spp. Conventional PCR for the LipL32 gene of L. interogans and the BPSS0120 and btfc-orf18 genes of B. pseudomallei was used for molecular detection. The overall seroprevalence of leptospirosis and melioidosis was 2.69% (95% confidence interval (CI): 0.99-5.76%) and 14.35% (95% CI: 10.03-19.65%), respectively. Six samples that showed positive MAT results were also positive for IHA. The serovars of Leptospira were Ranarum (5/6), Shermani (6/6), and both (5/6). Conventional PCR for the LipL32 gene of Leptospira spp. was positive in 10.31% of the samples (95% CI: 5.56-13.51%). However, there were no positive results for BPSS0120 and btfc-orf18 in B. pseudomallei. Active infection was detected only for leptospirosis; however, it can be assumed that pathogen exposure occurred in this group of animals because immunity could be detected. The routes of infection and elimination pathways of both bacteria remain unclear, and the mechanism of protection in non-human primates needs to be elucidated in further studies. Moreover, this health issue should be considered to prevent human infections in monkeys and their environment.

Prevalence and risk factors of Campylobacter infection in goats in southern provinces of Thailand.

Campylobacter is a foodborne pathogen that could be isolated from various types of livestock, including small ruminant. The objectives of the current study were to investigate the prevalence of Campylobacter spp. in goats reared in the south of Thailand and identify the risk factors associated with Campylobacter infection of goat in Thailand. A total of 600 rectal swabs of goats from 45 farms in 6 provinces of Southern Thailand were collected during 2016-2020. To analyse the risk factors associated with Campylobacter infection in goat, data collection was conducted via structured questionnaires. Campylobacter isolation was performed by direct plating on modified charcoal cefoperazone deoxycholate agar (mCCDA). Polymerase chain reaction (PCR) was used to identify genus and species of the isolates. In total, 68 out of 600 rectal swab samples (11.33%; 95% CI 8.96 to 14.21%) collected from goats were positive for Campylobacter. The prevalence of Campylobacter spp. in the province level ranged from 0 to 31.48%. Interestingly, Campylobacter lanienae was the dominant species, followed by Campylobacter coli and Campylobacter jejuni. To investigate the risk factors associated with Campylobacter infection in goats, farm management factors, i.e. deworming, fencing around the house, housing design, number of animals on farm, use of antibiotics and vaccination, were included in logistic regression analysis. Statistical analysis revealed that the use of antibiotics and deworming were associated with Campylobacter infections in goats. The current study emphasises that the prudent use of antimicrobials and good agricultural practices should be applied in goat farming.

Theileria infection in bullfighting cattle in Thailand.

Background and Aim: An apicomplexan protozoan parasite, namely, Theileria, primarily causes theileriosis in cattle worldwide. The virulence of the disease has been neglected because of it's low pathogenicity. However, the disease can have a substantial effect, depending on the virulence of the species, low host immunity, and coinfection. In Thailand, the molecular detection of Theileria infection in bullfighting cattle and its hematological alterations have not been reported. Thus, this study aimed to identify Theileria species in bullfighting cattle in Thailand. Materials and Methods: Blood samples were collected from bullfighting cattle presented at the Prince of Songkla University Animal Hospital and were determined on the basis of hematological evaluation and DNA extraction. Molecular detection using the 18s rRNA and merozoite surface antigen genes was conducted for Theileria spp. and Theileria orientalis, respectively. In addition, bidirectional sequencing of the positive samples was performed. Hematological alterations between Theileria infected and uninfected groups were statistically evaluated. Results: The levels of Theileria spp. and T. orientalis infection in bullfighting cattle were 44.62% (58/130) and 41.54% (54/130), respectively. Theileria orientalis, Theileria sinensis, and Theileria spp. infections were identified in bullfighting cattle samples. Hematological evaluation indicated that the red blood cell (RBC) level was significantly lower in Theileria-infected cattle. Conclusion: This study was the first to use molecular techniques in the identification of Theileria infection in bullfighting cattle in Thailand, with nearly one-half of the study population infected. Theileria infection in bullfighting cattle altered the RBC level, resulting in anemia. Therefore, tick control measures should be promoted.

First Molecular Detection of Babesia gibsoni in Stray Dogs from Thailand.

In southern Thailand, the increasingly growing population of stray dogs is a concern to public health and environmental safety because of the lack of medical attention and control. More importantly, these animals are considered reservoirs for many zoonotic pathogens. The objective of this study was to molecularly detect canine vector-borne pathogens, and to perform genetic characterization of Babesia gibsoni present in stray dogs from southern Thailand. Blood samples were collected from 174 stray dogs in two provinces (Songkhla and Narathiwat) in southern Thailand. PCR analyses were executed using specific primers based on the Babesia spp. 18S rRNA gene, Babesia gibsoni Internal transcribed spacer 1 (ITS1) region, Ehrlichia canis citrate synthase (gltA) gene, Hepatozoon spp. 18S rRNA gene and Anaplasma platys heat shock protein (groEL) gene. The most common canine vector-borne pathogen found infecting stray dogs in this study was Hepatozoon canis (24.7%) followed by A. platys (14.9%), Babesia vogeli (8.0%), B. gibsoni (6.3%), and E. canis (1.72%). Concurrent infection with more than one pathogen occurred in 72 cases. Phylogenetic analysis based on the ITS1 region and 18S rRNA gene revealed that the B. gibsoni isolates from this study shared a large proportion of their identities with each other and with other reported B. gibsoni genotypes from Asia. This study highlights the molecular detection of B. gibsoni in dogs in Thailand for the first time and presents the genetic characterization by sequencing the ITS1 region and 18S rRNA gene of B. gibsoni from Thailand. Follow-up studies are needed to elucidate the origin, distribution, and vectors of B. gibsoni parasites circulating in dogs in Thailand, as well as to determine to what extent dogs are important reservoir hosts for zoonotic canine vector-borne disease infection in the studied area.

Isolation and Characterization of Potential Salmonella Phages Targeting Multidrug-Resistant and Major Serovars of Salmonella Derived From Broiler Production Chain in Thailand.

Salmonella is a major foodborne pathogen that causes foodborne disease in humans through consumption of contaminated foods, especially those of animal origin. Multiple Salmonella strains are antibiotic-resistant due to the common use of antibiotics in farm animals, including broiler farms. In this study, an alternative strategy using phage-based treatment was evaluated against Salmonella isolated from the broiler production. The prevalence of Salmonella spp. showed up to 46.2 and 44.4% in bedding samples from the broiler farms located in eastern and southern Thailand, respectively. Overall, 21 samples (36.2%) were positive for Salmonella and eight serovars were recovered from cloacal swabs, bedding materials (rice husk), and boot swabs collected from five farms. Up to 20 Salmonella phages were isolated from seven water samples from wastewater treatment ponds, a river, and a natural reservoir in Songkhla province. Isolated phages were investigated, as well as their lysis ability on eight target Salmonella serovars derived from broiler farms, five foodborne outbreak-related serovars, and 10 multidrug-resistant (MDR) serovars. All phages showed a strong lytic ability against five serovars of Salmonella derived from broiler farms including Kentucky, Saintpaul, Schwarzengrund, Corvalis, and Typhimurium; three foodborne outbreak serovars including Enteritidis, Typhimurium, and Virchow; and eight MDR serovars including Agona, Albany, Give, Kentucky, Typhimurium, Schwarzengrund, Singapore, and Weltevreden. Three phages with the highest lysis potential including vB_SenS_WP109, vB_SenS_WP110, and vB_SenP_WP128 were selected for a phage cocktail preparation. Overall, a phage cocktail could reduce Salmonella counts by 2.2-2.8 log units at 6 h of treatment. Moreover, Salmonella did not develop a resistant pattern after being treated with a phage cocktail. Findings here suggest that a phage cocktail is an effective biocontrol to combat Salmonella derived from broiler production chain, other serovars linked to foodborne outbreaks, and MDR serovars.

Gastrointestinal parasite infections in fighting bulls in South Thailand.

BACKGROUND AND AIM: Bullfighting is booming in South Thailand, attracting tourists, and stimulating local economies. The bulls are well raised and practiced, but in many cases, the owners lack knowledge and understanding of the prevention of animal diseases, including parasitic infections. This study aimed to determine the occurrence of gastrointestinal (GI) parasite infection in fighting bulls. MATERIALS AND METHODS: A total of 1501 fecal samples were collected from bulls aged 2-5 years visiting the animal hospital of Prince of Songkla University during 2016-2019. The formalin ethylether concentration method was used to detect GI parasites in feces. RESULTS: The overall rate of GI parasite infection was 94.27%. Rumen fluke eggs were detected in 97.17% of all infected animals, followed in prevalence by strongyles (26.29%), Eurytrema spp. (2.83%), Fasciola spp. (2.47%), Trichuris spp. (0.35%), and Moniezia spp. (0.14%). Two protozoan genera were identified, Balantidium coli (6.64%) and Eimeria spp. (3.53%). Coinfection was observed in 33.99%. The five most common coinfections were rumen fluke with strongyles (20.85%), B. coli (4.66%), Eimeria spp. (1.55%), Eurytrema spp. (1.34%), and Fasciola spp. (1.06%). CONCLUSION: In addition to high GI parasite infection rates, zoonotic parasites were observed. Therefore, it is recommended that farmers should follow good sanitation and prevention practices to control parasitic infections in bulls, and proper hygienic precautions should be taken by the owners. Implementation of deworming programs using appropriateanthelmintic drugs as well as rotation of anthelmintic drug that have different chemical agent to prevent further drug resistance should be considered. The promotion of bull health management is highly recommended to protect humans from zoonotic diseases.

Investigation of Trypanosoma evansi infection in bullfighting cattle in Southern Thailand.

BACKGROUND AND AIM: Trypanosoma evansi infection has been reported in Thai livestock such as beef and dairy cattle. However, there is little information on T. evansi infection in bullfighting cattle in Southern Thailand. The aim of this study was to investigate the infection of T. evansi in bullfighting cattle presented for health checks at the Animal Hospital, Faculty of Veterinary Science, Prince of Songkla University, Thailand. MATERIALS AND METHODS: Blood and serum samples were collected from 177 bullfighting cattle from April 2016 to February 2017 after bullfighting matches. Animal inspected showed signs of fever, weight loss, or exercise intolerance. Investigation of T. evansi infection was tested using polymerase chain reaction (PCR) with TBR primers and using indirect enzyme-linked immunosorbent assay with T. evansi crude antigen. RESULTS: The seroprevalence of T. evansi in bullfighting cattle was 22.60% (40/177). The PCR results detected no parasite DNA in this study. However, bullfighting cattle may serve as T. evansi reservoirs. CONCLUSION: Health checking procedures for T. evansi should be promoted for bullfighting events so that infected animals can be quarantined in the preparatory stages of such events.

Investigation of Leptospira Infection in Stray Animals in Songkhla, Thailand: Leptospirosis Risk Reduction in Human.

Leptospirosis is a zoonosis of public health concern in Thailand. Human leptospirosis presents severe illness and can be fatal due to pulmonary hemorrhage, kidney failure, or cardiac impairment. Infected animals show no clinical signs and play an important role in the Leptospira infection of humans and other hosts. The prevalence of leptospirosis in stray animals in Thailand is unknown. The aim of this study was to investigate the incidence of Leptospira infection in stray animals including dogs and cats in Songkhla province, Thailand. A total of 434 blood samples were collected from 370 stray dogs and 64 stray cats during a population control program from 2014 to 2018. Screening the serum samples using the latex agglutination test to detect antibodies against Leptospira interrogans showed that 29.26% (127/434) were positive. There were 120 positive samples for stray dogs and 7 positive samples for stray cats. The detection of positive samples by polymerase chain reaction specific to the LipL32 of L. interrogans showed 1.61% (7/434) were positive. Stray cats (5/64) showed a higher prevalence than stray dogs (2/370), which might be because they are more likely to come into contact with rodents in their habitat. Although the active infection detected was low, the seroprevalence was high. This result indicated that the stray animals might not have been infected at the time of sample collection, but that they had been infected in the past or were in a latent period of infection. Therefore, they might serve as a leptospirosis reservoir for domestic animals and humans present in the same environment. The results show that stray animals need health care, such as vaccination, surveillance, and treatment, when infected to prevent or reduce the risk of transmission to humans.

Direct pentaplex PCR assay: An adjunct panel for meat species identification in Asian food products.

A direct pentaplex PCR assay was developed for the identification of meat from sources other than those declared on the packaging. Species-specific primers were designed, based on the mitochondrial cytochrome oxidase I (COI) gene. The assay amplified specific DNA fragments from dog (230 bp), duck (283 bp), buffalo (363 bp), goat (396 bp), and sheep (477 bp). The proposed method is capable of identifying target species accurately and is reproducible, sensitive and robust for use with real-world foods and food products. In total, 26 of 117 meat and commercial food products tested were shown to contain DNA from species not declared on the label.