Marcelo Signorini

Impact of probiotic administration on the health and fecal microbiota of young calves: a meta-analysis of randomized controlled trials of lactic acid bacteria.

Before weaning, dairy calves are susceptible to many pathogens which can affect their subsequent performance. The use of lactic acid bacteria (LAB) has been identified as a tool to maintain the intestinal microbial balance and to prevent the establishment of opportunistic pathogenic bacterial populations. However, a consensus has not been reached as to whether probiotics may be effective in reducing the prevalence of gastrointestinal diseases in young calves. The aim of this meta-analysis was to assess the effect of probiotics on diarrhea incidence and the intestinal microbial balance. LAB supplementation has been shown to exert a protective effect and to reduce the incidence of diarrhea (relative risk, RR=0.437, 95% confidence interval (CI) 0.251-0.761). In the subanalysis, this protective effect of the probiotics against diarrhea was observed only in trials that used whole milk (RR=0.154, 95% CI 0.079-0.301) and trials that used multistrain inocula (RR=0.415, 95% CI 0.227-0.759). Probiotics did not improve the fecal characteristics (standardized mean difference, SMD=-0.4904, 95% CI -1.011-0.035) and were unable to change the LAB:coliforms ratio (SMD=0.016, 95% CI -0.701-0.733). Probiotics showed a beneficial impact on the LAB:coliforms ratio in the subanalysis that included trials that used whole milk (SMD=0.780, 95% CI 0.141-1.418) and monostrain inocula (SMD=0.990, 95% CI 0.340-1.641). The probability of significant effects (probiotic positive effect) in a new study was >0.70 for diarrhea and fecal consistency. Whole milk feeding improved the action of the probiotic effect on the incidence of diarrhea and LAB:coliforms ratio. The probability to find significant effects in the diarrhea frequency and LAB:coliforms ratio was higher (P>0.85) if the new studies were conducted using whole milk to feed calves. This paper defines the guidelines to standardize the experimental designs of future trials. LAB can be used as growth promoters in calves instead of antibiotics to counteract the negative effects of their widespread use.

Exposure assessment of mycotoxins in cow's milk in Argentina.

A stochastic simulation model was developed to carry out the first quantitative risk exposure assessment of the mycotoxin level in cows milk produced in Argentina. The prevalence and concentration of aflatoxin M1 (AFM1), deoxynivalenol (DON) and zearalenone (ZEA) were modeled at various stages through milk processes complying with Argentinean practices. Concentration of AFM1 (0.059ppb), DON (0.338ppb) and ZEA (0.125ppb) in dairy milk were estimated. The proportion of feed samples that exceeded the maximum level accepted by European regulations for AFB1, DON and ZEA were estimated at 25.07%, 0.0% and 8.9%, respectively. The percentage of milk samples that exceeded the maximum level accepted for AFB1 by the MERCOSUR (0.5ppb) and the European Union regulations (0.05ppb) were 0.81 and 32.65, respectively. The probability distribution of AFM1 concentration in milk was affected by the carry-over rate equations applied in the model. Mycotoxin levels in corn silage and concentrated feeds were the factors most correlated with mycotoxin concentrations in milk. Therefore, agricultural practices, crop management and feed production require prompt attention regarding mycotoxin issues.

Evaluation of two alternative techniques for counting mesophilic aerobic bacteria in raw milk

Abstract Traditional techniques of microbiological analysis do not meet the current needs of having immediate analytical results in order to be able to make decisions in the process of food manufacturing, specially in HACCP application systems concerned with quality assurance. The development of modern techniques is showing new alternatives but it is important that the advantages for each of these new methodologies be considered for each application. The objective of this paper was to evaluate the advantages of two microbiological alternative techniques (dry rehydratable film and direct epifluorescent filter technique), applied to enumerating total aerobic mesophilic micro-organisms in refrigerated raw milk supplied by dairy farms, compared to the reference technique (standard plate count). The two alternative techniques under study showed good correlation levels with the standard in plate technique. Dry rehydratable film technique had higher costs but this technique allows the analysis of twice the quantity of samples in the same unit of time compared to the standard plate technique. This compensates for the higher costs. The epifluorescent technique gave the results in shorter time but it is necessary to use sophisticated equipment and trained personnel. Therefore, it is preferred when having the appropriate infrastructure.

Quantitative risk assessment for verocytotoxigenic Escherichia coli in ground beef hamburgers in Argentina

A quantitative risk assessment was developed for verocytotoxigenic Escherichia coli (E. coli VTEC) following hamburger consumption. The assessment considers initial contamination levels, cross-contamination and decontamination events during the cattle slaughter process and the distribution, storage and consumption patterns in Argentina and in similar countries in Latin-American. The model predicted an infection risk of 8.12x10(-7), a probability of Hemolytic Uremic Syndrome (HUS) of 4.6x10(-8) and a probability of mortality of 5.9x10(-9) per meal for adults. For children, the estimates per meal were 3.23x10(-7), 1.8x10(-8) and 6.31x10(-10) for infection, HUS and mortality, respectively. The risk of infection and HUS, were sensitive to the type of storage at home (r=-0.416), slaughterhouse storage temperature (r=0.240) and bacterial concentration in the cattle hide (r=0.239). There was an association between home preparation of hamburgers (r=-0.116) and the risk of illness, although this was a result of the type of storage at retail (r=-0.110) and at home and not their intrinsic characteristics. The most sensitive stages of the process were identified through the risk assessment and these can be used as a basis for measures of risk management.

Enzyme-linked immunosorbent assay to diagnose human leptospirosis: a meta-analysis of the published literature

We report an evaluation of the accuracy of ELISA for the detection of Leptospira-specific antibodies in humans. Eighty-eight studies published in 35 articles met all inclusion criteria and were submitted to meta-analysis. Pooled sensitivity and specificity were 0·779 (95% CI 0·770-0·789) and 0·913 (95% CI 0·908-0·917), respectively, and the area under the curve was 0·964. Heterogeneity across studies was statistically significant, but none of the sources of heterogeneity (disease stage, antigen used, antibody detected) could fully explain this finding. Although the convalescent stage of disease was significantly associated with higher diagnostic accuracy, IgM ELISA was the best choice, regardless of the stage of disease. Negative ELISAs (IgG or IgM) applied in the acute phase do not rule out leptospirosis due to the possibility of false-negative results. In this case it is advisable to request a second blood sample or to apply a direct method for leptospiral DNA.

Risk factors associated with failures of Varroa treatments in honey bee colonies without broodless period

The treatment against Varroa destructor has become a basic tool in beekeeping practices, mainly during autumn. The treatment effectiveness should be improved by identifying variables affecting the final outcome. The aim of this study was to identify the risk factors associated with the treatment outcome achieved during autumn control of Varroa destructor. The mite infestation after treatment was evaluated in 62 apiaries and data regarding management practices were collected by means of a questionnaire. A mixed-effects model was constructed to associate management variables with the risk of treatment failure occurrence. Colonies with high mite levels prior to treatment (P = 0.002) and owned by beekeepers who did not frequently replace queens (P = 0.001) were associated with a higher risk of treatment failure. Other beekeeping practices indirectly improved treatment effectiveness. An integrated strategy for controlling mites that includes chemotherapy and suitable beekeepers management is needed to keep mite populations low during winter.

Risk factors associated with the presence of Varroa destructor in honey bee colonies from east-central Argentina

Varroa destructor is considered one of the major threats for worldwide apiculture. Damage caused by varroa mite includes body weight loss, malformation and weakening of the bees. It was also suggested as the main cause associated with colony winter mortality and as an important vector for several honey bee viruses. Little is known about multiple factors and their interaction affecting V. destructor prevalence in apiaries from South America. The aim of this study was to identify risk factors associated with V. destructor prevalence in east-central Argentina. Parasitic mite infestation level and colony strength measures were evaluated in 63 apiaries distributed in 4 different regions in east-central Argentina in a cross sectional study. Data regarding management practices in each apiary were collected by means of a questionnaire. A mixed-effects logistic regression model was constructed to associate management variables with the risk of achieving mite infestation higher than 3%. Colonies owned by beekeepers who indicated that they did not monitor colonies after mite treatment (OR=2.305; 95% CI: 0.944-5.629) nor disinfect hives woodenware material (OR=2.722; 95% CI: 1.380-5.565) were associated with an increased risk of presenting high intensity infestation with V. destructor (>3%). On the other hand, beekeepers who reported replacing more than 50% of the queens in their operation (OR=0.305; 95% CI: 0.107-0.872), feeding colonies protein substitute containing natural pollen (OR=0.348; 95% CI: 0.129-0.941) and feeding colonies High Fructose Corn Syrup (HFCS) (OR=0.108; 95% CI: 0.032-0.364), had colonies that were less likely to have V. destructor infestations above 3%, than beekeepers who did not report using these management practices. Further research should be conducted considering that certain management practices were associated to mite infestation level in order to improve the sanitary condition in the colonies. Epidemiological studies provide key information to design surveillance programs against one the major threat to worldwide beekeeping.

Quantitative risk assessment of human campylobacteriosis by consumption of salad cross-contaminated with thermophilic Campylobacter spp. from broiler meat in Argentina

Here, we developed a quantitative risk assessment for thermophilic Campylobacter spp. related to the consumption of salad prepared alongside broiler meat. The assessment considered initial contamination levels, cross-contamination and decontamination events during the broiler slaughter process and distribution, and storage and consumption patterns in Argentina and other Latin American countries. The model predicted an infection risk of 3.32×10(-4) per serving. This estimation was variable according to the dose-response model used. Considering the number of chickens slaughtered annually in Argentina, the estimated number of people who could suffer campylobacteriosis related to poultry meat consumption was, on average, 484,304. The risk of human campylobacteriosis was most sensitive to the probability of infection from a Campylobacter (r=0.72), the number of Campylobacter spp. per serving (r=0.40), the frequency of washing the cutting board (r=-0.31), the preparation of raw poultry before salad using the same cutting board (r=0.14), and the frequency of hand washing (r=-0.14). The most sensitive stages of the process identified through the risk assessment can be used as a basis for measures of risk management. Public campaigns on hygiene habits during food preparation at home should focus on the importance of washing the cutting board before preparing raw and ready-to-eat foods and of washing the hands during food preparation.

Occurrence of thermotolerant Campylobacter spp. at different stages of the poultry meat supply chain in Argentina

AIMS: The objectives of this study were to investigate the occurrence and concentration of thermotolerant Campylobacter spp. at different stages of the poultry meat supply chain in Argentina. METHODS: Three integrated poultry companies were sampled. Each supply chain was considered at different stages from the reproductive farm to chicken meat at a retail market. The stages sampled were: (a) hens from breeder flocks, (b) eggs in the incubator, (c) broiler chickens in flocks (aged <1 week and >5 weeks), (d) chickens at a slaughterhouse, and (e) chicken meat at a retail market. The chickens sampled along each supply chain were in the same batch. Samples collected were: (a) cloacal samples from hens and chickens on the farms, (b) fertile eggs, (c) feed, water and litter from flocks, (d) chicken carcasses from the slaughterhouse and retail market, and (e) caeca and livers from the slaughterhouse. Samples obtained were examined for Campylobacter spp. The isolates were biotyped and the genus and species identified by PCR. Campylobacter spp. on chicken carcasses at slaughterhouse and retail market were enumerated. RESULTS: The highest proportions of Campylobacter positive samples were observed in carcasses at retail (25/30, 83.3%) and faecal samples from breeding hens (27/45, 60.0%). Only 3.3% (3/90) samples collected from broiler chickens aged <1 week were positive, but the percentage of positive samples had risen to 28.9% (26/90) by the end of the rearing period. The proportions of Campylobacter positive carcasses and caecal contents at the slaughterhouse were both 33.3% (10 of 30 samples each). The concentration of Campylobacter contamination observed on carcasses at retail markets ranged from no bacteria/carcass to 3.71 log10 cfu/carcass. CONCLUSIONS: The data obtained provide essential information for future quantitative risk assessments aiming to estimate the probability of a person contracting campylobacteriosis following consumption of broiler meat in Argentina. CLINICAL RELEVANCE: The proportions of Campylobacter-positive samples found in this preliminary study indicate that a large proportion of the cases of human gastroenteritis in Argentina may be due to this pathogen. Human cases of gastroenteritis should be studied in greater detail and measures should be developed to reduce the proportion of poultry products that are contaminated by Campylobacter species.

Zoonoses in rural veterinarians in the central region of Argentina

OBJECTIVE To estimate the frequency of zoonoses in rural veterinarians and to search for risk factors. DESIGN Cross-sectional study based on an anonymously answered structured questionnaire. SETTING The interviewees participated in mandatory continuing education classes scheduled throughout the province by the College of Veterinary Surgeons. PARTICIPANTS Overall, 741 professionals were surveyed, and 75.8% (n = 562) of them completed the structured questionnaire. MAIN OUTCOME MEASURES Cumulative incidence (CIR) and incidence density (IDR) rates, standardised rates, χ(2) , Students t-test, Pearsons correlation coefficient and logistic regression. RESULTS The CIR for all zoonoses was 34.1% (brucellosis, 29.1%; toxoplasmosis, 2.1%; leptospirosis, 0.6%; tuberculosis, 0.6%; anthrax, 0.6%; ringworm, 0.4%; other, 0.6%). The IDR for the period 1964-2008 was estimated to be 20.7% (19.5% for brucellosis). The brucellosis IDR decreased between 1964 and 2008 and was higher during early post-graduation. The risk of brucellosis was associated with the number of years of practice and the geographical area. Sixty-nine respondents had at least one day of absence from work (24.0 ± 27.8 days). CONCLUSIONS A high frequency of zoonoses was reported by veterinarians with a large animal practice. Although the rate of zoonoses may be decreasing, further studies are needed to confirm this finding. A joint effort of all institutions is needed to prevent zoonoses among private practitioners.