Occurrence and seasonal variation of aflatoxin M1 in raw cow milk collected from different regions of Algeria.

BACKGROUND AND AIM: Aflatoxins are metabolites of molds that exert potentially toxic effect on animals and humans. This study aimed to investigate the occurrence of aflatoxin M1 (AFM1) in raw cow milk collected during 1 year (2016-2017) from different regions of Algeria and risk factors associated with the contamination. MATERIALS AND METHODS: During the survey period, 84 samples of raw milk were collected in three regions of Algeria (northeast, north center, and northwest) during four seasons. AFM1 levels were analyzed by competitive enzyme-linked immunosorbent assay. RESULTS: AFM1 was detected in 39 (46.43%) samples (total mean concentration, 71.92 ng/L; range, 95.59-557.22 ng/L). However, the AFM1 levels exceeded the maximum tolerance limit set by the Food and Drug Administration in the USA (500 ng/L) in only 1 sample (1.19%). Statistical analysis revealed significant differences (p˂0.005) between AFM1 levels in milk samples collected in the spring and autumn. The mean AFM1 levels in samples collected in the spring were significantly higher than those in samples collected in autumn. CONCLUSION: The survey indicates that farmers involved in milk production should be made aware of the adverse effects of aflatoxin contamination in animal feed. A systematic control program of supplementary feedstuff for lactating cows should be introduced by the public health authorities.

Understanding the epidemiology of sheep-pox outbreaks among vaccinated Algerian sheep and post vaccination evaluation of the antibodies kinetics of the commercially used vaccine.

Sheep pox is a disease of veterinary concern to small ruminant producers and veterinary diagnosticians, because of the associated tangible economic losses. The epidemiological analysis of sheep pox, among vaccinated sheep flock in Algeria from 2007 to 2016, showed that the disease outbreaks occurred every year and across all Algeria region with an average of 44.9 outbreaks per year, these outbreaks correlate with the region climate, the flocks' density and the transhumance practices. The one-year post vaccination antibody kinetics evaluation study of the commercially used vaccine in Algeria demonstrated a mild humoral response, the neutralization index range between 0.73 and 1.22. Therefore, the present study recommends a challenge study, using a virulent local strain, to evaluate the vaccine efficacy. Furthermore, quality control approach for the vaccine production processes is required.

Detection of multidrug resistant Escherichia coli in the ovaries of healthy broiler breeders with emphasis on extended-spectrum ß-lactamases producers.

In the last few years, antimicrobial resistant (AMR) Escherichia coli have been detected in newborn chickens suggesting their vertical transmission from breeding birds to their offspring. However, little is known about the presence of AMR E. coli in the reproductive organs of broiler breeders. The aim of this study was to investigate the presence of E. coli in the ovaries of healthy broiler breeders and to study their antimicrobial resistance. Samples from broiler breeders (n = 80) collected from 80 different broiler breeder flocks were included in this study. Antibiotic susceptibility testing was performed using disk diffusion method according to Clinical and Laboratory Standards Institute guidelines. Minimal inhibitory concentrations (MICs) of five antimicrobial agents were determined by Etest. PCR and sequencing were used to detect the blaESBL genes. E. coli were detected in the ovaries of thirty seven out of 80 (46.25%) sampled flocks. High levels of resistance to various first-line antimicrobial agents were recorded in E. coli isolates. This study showed that 89.18% of E. coli isolates were multidrug resistant (MDR). Furthermore, MDR extended-spectrum ß-lactamases (ESBL)-producing E. coli were detected in the ovaries of four different broiler breeder flocks. Molecular characterization revealed that three isolates harboured blaCTX-M-1 gene and one isolate expressed blaSHV-12 gene. In addition, one blaCTX-M-1 -producing E. coli co-harboured the blaTEM-1 gene. These findings would contribute to a better epidemiological understanding of MDR E. coli for improve existing preventive strategies in order to reduce the dissemination of antimicrobial resistance in the broiler production system.

Epidemiology of dog-mediated zoonotic diseases in Algeria: a One Health control approach.

Globally, human-dog interactions cause significant social, economic and human health costs. Public health problems linked with dogs include mainly zoonotic diseases. Recently the concept of a global and integrative approach to improve the health and well-being of people, animals and the environment has been strongly endorsed. This concept, often referred to as One Health, also reflects the collaboration in the field of surveillance and monitoring. Because humans and animals often suffer from the same pathogens and share the same environment, a cross-sector approach integrating human and animal disease surveillance information is required. The aims of the present study were to describe the incidence of dog-mediated zoonotic diseases in Algeria between 2010 and 2017, and to propose a One Health approach to control these diseases in Algeria. Rabies, leishmaniasis and echinococcosis are the major zoonoses in Algeria, with a reported average number of deaths per year, respectively, of 18, 7947 and 387. These zoonoses occur with the uncontrolled proliferation of household waste deposits, particularly in and around urban area which maintain the presence of stray dogs. The persistence of these diseases indicates the need for greater partnership and collaboration among multiple sectors, including medical doctors, veterinarians, ecologists, environmentalists and law-enforcement agents. Such partnerships permit the sharing of information, facilities and resources under a One Health approach; permit rapid communication among disciplines as well as interdisciplinary training/education opportunities and raising awareness among human population; and allow a combined effort towards disease surveillance/control, which will consequently improve the efficiency of the control programmes.

Antibacterial activity of Thymus vulgaris essential oil alone and in combination with cefotaxime against blaESBL producing multidrug resistant Enterobacteriaceae isolates.

The aim was to evaluate the susceptibility of blaESBL producing Enterobacteriaceae to Slovakian Thymus vulgaris essential oil (TVEO) alone and in combination with cefotaxime (CTX). TVEO composition was determined by gas chromatograph-mass spectrometer (GC/MS). Susceptibility to 21 antibiotics was determined by disc diffusion assay. Genes characterization for resistance to ß-lactams was accomplished by polymerase chain reaction (PCR). The antibacterial activity was investigated by standard methods. The synergistic interaction was determined by checkerboard test. Thymol (34.5%), p-cymene (22.27%) and linalool (5.35%) were the major components present in the TVEO. The identified strains were multi-drug resistant (MDR). TVEO showed high activity against all MDR strains, including blaESBL producing isolates, with inhibition zones and MIC values in the range of 24-40 mm/10µL and 2.87-11.5 µg/mL, respectively. TVEO in combination with CTX showed a synergistic action against blaSHV-12 producing Escherichia coli (FICI 0.28) and an additive effect vs ESBL producing Enterobacter cloacae (FICI 0.987).

Isolation of Escherichia coli carrying the blaCTX-M-1 and qnrS1 genes from reproductive organs of broiler breeders and internal contents of hatching eggs.

This study aimed to characterize two third-generation cephalosporins- and quinolone-resistant Escherichia coli (TGCs- and Q-R-Ec) isolates recovered from the ovaries of a broiler breeder flock and the internal contents of hatching eggs produced by the broiler breeder flock. Clonal relatedness was determined by multilocus sequence typing (MLST). The isolates displayed the same multidrug resistance profile, with resistance to ampicillin, ticarcillin, piperacillin, cefazollin, cephalothin, cefotaxime, nalidixic acid, tetracycline and sulfonamides. Double disk synergy test demonstrated that the two isolates presented an ESBL phenotype. PCR and sequencing results showed that both the isolates harbored the blaCTX-M-1 and qnrS1 genes. MLST revealed a novel allele combination, designated as ST461, in these isolates. This study would contribute to the molecular epidemiological understanding of TGCs- and/or Q-R-Ec.

The epidemiology of dog rabies in Algeria: Retrospective national study of dog rabies cases, determination of vaccination coverage and immune response evaluation of three commercial used vaccines.

Rabies is a neglected zoonotic disease. Several animal species can transmit rabies, but domestic dogs are the main reservoir implicated in rabies transmission to human and other susceptible animals. There is a scarcity of data regarding canine rabies in Algeria. Hence, this report is the first study that attempts to understand the epidemiology of dog rabies through a retrospective national study of rabies cases between 2010 and 2016, determination of canine vaccination coverage and evaluation of immune response of three rabies commercial vaccines used in Algeria. Our results revealed the persistent nature of dog rabies in Algeria; with a mean of 131 positive cases per year. The vaccination coverage findings indicated that 30.81% (126/409) of the reported vaccinated dog and 47.98% (261/544) of all studied dogs presented low level of rabies antibodies titres (less than 0.5 IU), and were therefore considered not immune to rabies. Additionally, our vaccine evaluation findigs showed a weak immune response of inactivated vaccine without adjuvant. Even with adjuvanted inactivated and attenuated live vaccines, several vaccinated dogs did not develop vaccine protection. Therefore, our results recommend further studies to test the efficiency of adding a booster vaccine shot, especially in primo-vaccinated dogs to optimize the vaccination protocol; rabies is potentially fatal zoonosis, tolerating no vaccine failure. Moreover, the present study establish the necessity to re-examine the national rabies control program, implemented in 1996, particularly concerning population education, vaccination strategy, surveillance and campaigns monitoring.

Characterization of quinolone-resistant Enterobacteriaceae strains isolated from poultry in Western Algeria: First report of qnrS in an Enterobacter cloacae.

AIM: Multidrug-resistant (MDR) Enterobacteriaceae have frequently been reported, in both human and veterinary medicine, from different parts of the world as a consequence of antibiotic usage. However, there is a lack of published data regarding antimicrobial resistance in non-Escherichia coli (E. coli) Enterobacteriaceae from animals in Algeria. This study aimed to evaluate the frequency of resistance to antibiotics with a focus on quinolones and to investigate the presence of qnr genes inEnterobacteriaceaeof poultry origin. MATERIALS AND METHODS: A total of 310 samples of poultry origin were collected from 2010 to 2014 from broiler and layer farms and hatcheries located in different geographic areas of Western Algeria (including Mostaganem, Oran, Mascara, Relizane, Chlef, Tiaret, and Tissemsilt). Antimicrobial susceptibility testing was performed using disc diffusion assay. Polymerase chain reaction and sequencing accomplished the characterization of qnr genes (qnrA, qnrB, and qnrS). RESULTS: A total of 253 Enterobacteriaceaestrains were isolated in this study. These isolates exhibited high levels of resistance to quinolones and other families of antibiotics. All the strains isolated in this study were resistant to at least one antibiotic. Among them, 233 (92.09%) were considered MDR. Among the 18 randomly selected nalidixic acid (NA)-resistant Enterobacteriaceaeisolates, one E. coli and one Enterobacter cloacae were carrying qnrS1. By contrast, qnrA and qnrB were not detected in this study. CONCLUSION: This is the first report on the identification of the qnrS gene in E. cloacae isolated from animal source in Algeria. Further studies have to be conducted to determine the real prevalence of qnr genes.

Seroprevalence of West Nile virus antibodies in equids in the North-East of Algeria and detection of virus circulation in 2014.

West Nile fever (WNF) is a viral disease of wild birds transmitted by mosquitoes. Humans and equids can also be affected and suffer from meningoencephalitis. In Algeria, since the 1994 epidemic, no data on WNV circulation was available until 2012. In September 2012, a fatal human case of WNV neuro-invasive infection occurred in Jijel province. This study describes the first seroprevalence study of West Nile virus (WNV) antibodies conducted in the equine population in Algeria. During 2014, serum samples were collected from 293 equids (222 donkeys and 71 horses) asymptomatic and unvaccinated for WNV in three localities in Northeastern wetlands of Algeria. Antibodies against WNV were found in 51 samples (seroprevalence 17.4%) of sampled equids, distributed as follows: 19 (seroprevalence 26.8%) horses and 32 (seroprevalence 14.4%) donkeys. Moreover 7 horses coming from Blida, in the center of Algeria, were tested before and after an 8-months stay in North-East Algeria. We observe a seroconversion in 2 horses, showing WNV circulation in 2014 in this specific region of Algeria.

Abortion and various associated risk factors in small ruminants in Algeria.

Identification of the causes of abortion among the huge population of small ruminants in Algeria (≈31 millions heads), is an important task for the control of livestock productivity and viability scourges to the small ruminants industry. Optimal production and utilization is constrained by a number of factors: disease, poor feeding and low management skills. Therefore, in the present study the prevalence of abortion in Algerian small ruminant's flocks was estimated and its possible association was correlated with infectious (PPR, BT and Brucellosis seropositivity) and managerial (flock size, grazing system, type of farming, and contact with other flocks) risk factors. The present study showed an overall flock prevalence of small ruminant's abortion as 75.33% (113/150) [95% CI 71.72-78.94%]. The risk factor analysis using multivariable logistic regression recognized the north-western and the steppe region as well as PPR positivity as a risk factor for abortion in Algerian small ruminant's flocks. The odds of flock abortion was 11.47 [95% CI 2.39-54.88; P=0.002] and 10.31 [95% CI 1.28-82.88; P=0.028] times higher in north-western and steppe regions respectively compared to other region. Also the presence of PPRV infection in small ruminant flocks amplified the odds by 6 times [95% CI 2.221-17.427; P=0.001].Surprisingly, the univariate analysis for the other risk factors associated with abortions in Algerian small ruminant flocks indicated no statistically significant links with bluetongue (P=1.000) and brucellosis seropositivity (P=0.334). Flock size (P=0.574), type of farming (P=0.443), grazing system (P=0.117) and contact with other flocks (P=0.245) was also not statistically significant. Our results revealed that abortion in small ruminants is a challenge to farmers and PPR was chiefly linked to it. Therefore an effective vaccination and control programme is advocated for small ruminants in Algeria.

Seroprevalence, distribution and risk factor for peste des petits ruminants (PPR) in Algeria.

Peste des petit ruminants (PPR) is a highly contagious and infectious viral disease of small ruminants with severe socio-economic implications. The disease was first reported in the Southern part of Algeria in 2011 and by February 2012 it has reached the central part of the country. Estimating national prevalence, distribution and identification of risk factors remains a key component in understanding the epidemiology and control of the disease. The present study was carried out between January and June 2014, to include a detailed description of flock and within-flock seroprevalence and risk association between PPR seropositivity and various flock management factors in Algeria. A total of 150 flocks randomly sampled across the country were investigated and 4552 serum samples were collected from 3336 sheep and 1216 goats, respectively. C-ELISA was used to detect the presence of antibodies in small ruminant animals as an indicator of PPRV exposure. The results showed an overall true flock seroprevalence of 30.45% [95% CI 23.76-37.14] with a mean of the true within-flock prevalence as 29.87%±2.11. The mean of the true within-flock prevalence in mixed flocks (12.93%±1.85) was however found to be significantly higher than sheep flocks (5.74%±1.06). Also the mean of the true within-flock prevalence was found to be significantly higher in adult (35.36%±3.13) compared to young animals (21.83%±2.47) and in females (33.11%±2.87) compared to males (22.14%±2.31). The univariate analysis revealed that PPR overall flock seroprevalence was significantly higher (P<0.20) in large flock (50.61%) than in small flock (33.33%), in mixed flock (56.7%) than in sheep flock (35.35%) and in the flocks that had contact with other flocks (46.5%) compared to those who had not (30.6%). However the differences among studied regions and grazing system were not statistically significant. For the risk factor analysis, univariate analysis of variables followed by a multiple logistic regression identified mixed flocks [OR=2.64, 95% CI 1.30-5.38; P=0.007] and contact with other flocks [OR=2.27, 95% CI 0.99-5.21; P=0.053] as risk factors in the spread of the disease. In conclusion, this study revealed a high seroprevalence of PPR in Algerian small ruminants, therefore the establishment of early warning systems and comprehensive implementation of control measures are advocated to improve animal welfare and reduce economic losses.

First serological and molecular evidence of PPRV occurrence in Ghardaïa district, center of Algeria.

In February 2012, an outbreak of peste des petits ruminants (PPR) was suspected in Ghardaïa district at the center of Algeria. Clinical, serological, and molecular investigations were performed to confirm the occurrence of PPRV. The overall morbidity, mortality, and case fatality rates of the ten flocks investigated were 12.2, 2.5, and 20.3 %, respectively. At the flock level, positivity to PPR was 100, 90, and 100 % by competitive ELISA (c-ELISA), RT-PCR of blood samples, and oculo-nasal swabs, respectively. At the individual levels, the present study showed that out of 186 samples collected from the same animals 17/62 (27.41 %), 14/62 (22.85 %), and 36/62 (58.06 %) were positive by c-ELISA, RT-PCR of blood samples, and RT PCR of oculo-nasal swabs, respectively. The positivity of PPR was significantly higher using RT-PCR of oculo-nasal swabs than c-ELISA and RT-PCR of blood samples. The N gene partial sequence of five PPRV-positive amplicons revealed 100 % homology among them and phylogenetically belonged to lineage IV. The sequences also showed similarity range of 97-99 % with the strains implicated in the Moroccan and Tunisian outbreaks, however, suggesting that a similar strain is circulating across this area of the Maghreb and highlighting the need for a regional control approach.