M.R. Rosmini

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.

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.

Molecular microbial analysis of Lactobacillus strains isolated from the gut of calves for potential probiotic use.

The intestinal microbiota has an influence on the growth and health status of the hosts. This is of particular interest in animals reared using intensive farming practices. Hence, it is necessary to know more about complexity of the beneficial intestinal microbiota. The use of molecular methods has revolutionized microbial identification by improving its quality and effectiveness. The specific aim of the study was to analyze predominant species of Lactobacillus in intestinal microbial ecosystem of young calves. Forty-two lactic acid bacteria (LAB) isolated from intestinal tract of young calves were characterized by: Amplified Ribosomal DNA Restriction Analysis (ARDRA), by using Hae III, Msp I, and Hinf I restriction enzymes, and 16S rDNA gene sequencing. ARDRA screening revealed nine unique patterns among 42 isolates, with the same pattern for 29 of the isolates. Gene fragments of 16S rDNA of 19 strains representing different patterns were sequenced to confirm the identification of these species. These results confirmed that ARDRA is a good tool for identification and discrimination of bacterial species isolated from complex ecosystem and between closely related groups. This paper provides information about the LAB species predominant in intestinal tract of young calves that could provide beneficial effects when administered as probiotic.

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.

Probiotics and broiler growth performance: a meta-analysis of randomised controlled trials

Abstract 1. The aim of this meta-analysis was to investigate the effects of probiotics on the growth performance of broilers. PubMed, Scopus and Scholar Google databases were searched in all languages from 1980 to 2012. The studies in the meta-analysis were only selected if they were randomised and controlled experiments using broilers without apparent disease and the results were published in peer-reviewed journals. 2. A total of 48 and 46 studies were included to assess probiotic effects on body weight gain (BWG) and feed efficiency (FE), respectively. Probiotics increased BWG compared to controls (SMD = 0.661, 95% CI 0.499 to 0.822) and improved FE (SMD = − 0.281, 95% CI −0.404 to −0.157) in the pooled standardised mean difference random effect model, considering the source of heterogeneity and publication biases. However, there are evidences of publication bias and heterogeneity, so the results of this meta-analysis should be considered with caution. Applying the Duval and Tweedie’s trim-and-fill methods, the adjusted value for BWG was 0.0594 (95% CI −0.122 to 0.242), and the adjusted value for FE did not show any modifications. 3. The meta-analysis showed that application of probiotics via water resulted in greater BGW and FE than administration through the feed. The effect was not related to the use of mono-strain or multi-strain probiotics, although it may depend on the strain used. The number of broilers and the duration of the experiments had an impact on the outcomes. 4. Additional studies should be conducted with the aim to identify the covariates which can explain the differences in the estimated effect sizes.

Pathogen translocation and histopathological lesions in an experimental model of Salmonella Dublin infection in calves receiving lactic acid bacteria and lactose supplements.

The purpose of this study was to evaluate the capacity of a lactic acid bacteria (LAB) inoculum to protect calves with or without lactose supplements against Salmonella Dublin infection by evaluating histopathological lesions and pathogen translocation. Fifteen calves were divided into three groups [control group (C-G), a group inoculated with LAB (LAB-G), and a group inoculated with LAB and given lactose supplements (L-LAB-G)] with five, six, and four animals, respectively. The inoculum, composed of Lactobacillus (L.) casei DSPV 318T, L. salivarius DSPV 315T, and Pediococcus acidilactici DSPV 006T, was administered with milk replacer. The LAB-G and L-LAB-G received a daily dose of 109 CFU/kg body weight of each strain throughout the experiment. Lactose was provided to the L-LAB-G in doses of 100 g/day. Salmonella Dublin (2 × 1010 CFU) was orally administered to all animals on day 11 of the experiment. The microscopic lesion index values in target organs were 83%, 70%, and 64.3% (p < 0.05) for the C-G, LAB-G, and L-LAB-G, respectively. Administration of the probiotic inoculum was not fully effective against infection caused by Salmonella. Although probiotic treatment was unable to delay the arrival of pathogen to target organs, it was evident that the inoculum altered the response of animals against pathogen infection.

Probióticos en pollos parrilleros: una estrategia para los modelos productivos intensivos ☆

The broiler industry has become an important economic activity in Argentina. Global production of broiler meat has been growing in Argentina faster than for any other meats, possibly due to declining poultry prices and increasing incomes. Modern rearing systems can produce broilers ready to slaughter in 50 days, with the required 2.7kg of weight and a feed conversion of about 1.6kg feed/kg of meat. Nevertheless, broilers raised under these intensive conditions are exposed to various stressors every day. For many years, feed supplementation with antibiotics was widely used to stabilize the gut flora, improve general parameters and prevent avian diseases. However, the utility of antibiotics has been questioned because of the emergence and spread of antibiotic-resistant bacteria in meat. Therefore, there is a renewed interest in finding viable alternatives to antibiotics. One potential method is the supplementation of broiler diets with probiotics. This review provides an updated summary of the use of probiotics to improve sanitary conditions and enhance performance in broilers, demonstrating the role of probiotics as a reliable option to replace antimicrobial growth promoters.

Quantification of FITC-labelled probiotic Lactobacillus salivarius DSPV 001P during gastrointestinal transit in broilers

The knowledge related to the fate of probiotics in the complex environment of the intestinal microbiota in broilers is just beginning to be elucidated; however, it is not yet well understood. A good method to investigate the mechanisms by which probiotics mediate their effects is to mark probiotic bacteria and trace them. The aim of this research was to develop a new method to estimate in vivo fluorescein isothiocyanate (FITC)-labelled Lactobacillus salivarius DSPV 001P counts during passage through the gastrointestinal tract (GIT) of broilers. Forty-five, 1 d old Cobb broilers were used in this trial. Programmed necropsies were performed 30 min, 6 h, and 12 h after the administration of the probiotic bacterium, and samples of liver, crop, duodenum, caecum, and bursa of fabricius were collected. To determine the spatial and temporal transit of L. salivarius DSPV 001P in broilers, the number of bacteria as well as its respective fluorescent signal produced by FITC were measured. In order to observe the relationship between the variables, a logistic regression analysis was applied. The amount of fluorescence could be used as an indicator of fluorescent probiotic bacteria in the crop and duodenum 30 min after probiotic bacterium supplementation. In addition, the fluorescent signal could be used to estimate bacterial counts in caecum 6 and 12 h after L. salivarius DSPV 001P administration. To the best of our knowledge, this research is the first in vivo trial to employ the bacterial FITC-labelling technique in order to enumerate probiotic bacteria during gastrointestinal transit in broilers.

Quantitative Risk Assessment of Human Trichinellosis Caused by Consumption of Pork Meat Sausages in Argentina

In Argentina, there are three known species of genus Trichinella; however, Trichinella spiralis is most commonly associated with domestic pigs and it is recognized as the main cause of human trichinellosis by the consumption of products made with raw or insufficiently cooked pork meat. In some areas of Argentina, this disease is endemic and it is thus necessary to develop a more effective programme of prevention and control. Here, we developed a quantitative risk assessment of human trichinellosis following pork meat sausage consumption, which may be used to identify the stages with greater impact on the probability of acquiring the disease. The quantitative model was designed to describe the conditions in which the meat is produced, processed, transported, stored, sold and consumed in Argentina. The model predicted a risk of human trichinellosis of 4.88 × 10−6 and an estimated annual number of trichinellosis cases of 109. The risk of human trichinellosis was sensitive to the number of Trichinella larvae that effectively survived the storage period (r = 0.89), the average probability of infection (PPinf) (r = 0.44) and the storage time (Storage) (r = 0.08). This model allowed assessing the impact of different factors influencing the risk of acquiring trichinellosis. The model may thus help to select possible strategies to reduce the risk in the chain of by‐products of pork production.

Probiotic effect on calves infected with Salmonella Dublin: haematological parameters and serum biochemical profile

The aim of this study was to evaluate the effect of a probiotic/lactose inoculum on haematological and immunological parameters and renal and hepatic biochemical profiles before and during a Salmonella Dublin DSPV 595T challenge in young calves. Twenty eight calves, divided into a control and probiotic group were used. The probiotic group was supplemented with 100 g lactose/calf/d and 1010 cfu/calf/d of each strain of a probiotic inoculum composed of Lactobacillus casei DSPV318T, Lactobacillus salivarius DSPV315T and Pediococcus acidilactici DSPV006T throughout the experiment. The pathogen was administered on day 11 of the experiment, at an oral dose of 109 cfu/animal (LD50). Aspartate aminotransferase (AST), gamma glutamyl transpeptidase (GGT), urea, red blood cells, haemoglobin, haematocrit, mean cell haemoglobin (MCH), mean corpuscular volume, mean corpuscular haemoglobin concentration (MCHC), white blood cells, lymphocytes, neutrophils, band neutrophils, monocytes, eosinophils, basophils and the neutrophils/lymphocytes ratio were measured on days 1, 10, 20 and 27 of the experiment. In addition, animals were necropsied to evaluate immunoglobulin A (IgA) production in the jejunal mucosa. The most significant differences caused by the administration of the inoculum/lactose were found during the acute phase of Salmonella challenge (9 days after challenge), when a difference between groups in neutrophils/lymphocytes ratio were detected. These results suggest that the probiotic/lactose inoculum administration increases the calfs ability to respond to the disease increasing the systemic immune response specific. No differences were found in haemoglobin, haematocrit, MCH, MCHC, AST, urea, GGT, band neutrophils, eosinophils, monocytes and IgA in the jejunum between the two groups of calves under the experimental conditions of this study. Further studies must be conducted to evaluate different probiotic/pathogens doses and different sampling times, to achieve a greater understanding of the effects of this inoculum on intestinal infections in young calves and of its mechanism of action.