Identification of Lactobacillus spp. from broiler litter in Brazil

Identificação de Lactobacillus spp. de cama de frango no Brasil

VETERINARY MICROBIOLOGY

Identification of Lactobacillus spp. from broiler litter in Brazil

Identificação de Lactobacillus spp. de cama de frango no Brasil

Ronaldo S. Paço^I; Ivani Lucia Leme^II; José Américo Bottino^I; Antônio José Piantino Ferreira^I,*

^IDepartamento de Patologia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo, SP, Brasil

^IIDepartamento de Doenças Infecciosas e Parasitárias, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP, Brasil

ABSTRACT

Lactobacillus spp. were identified in 100 broiler litter samples collected from different poultry-rearing regions in Brazil. Ten different Lactobacillus species were identified: L. plantarum, L.casei subsp. pesudoplantarum, L. delbrueckii subsp. delbrueckii, L. reuteri, L. murinus, L. agilis, L. delbrueckii subsp. lactis, L. salivarus subsp. salicinus, L. viridenscens and L. amylophilus.

Key words:Lactobacillus spp., broiler litter, chicken.

RESUMO

Foram identificadas cepas de Lactobacillus spp. de 100 amostras de camas de frango coletadas de diferentes regiões de produção avícola do Brasil. Foram isoladas dez espécies diferentes de Lactobacillus: L. plantarum, L. casei subsp. pseudoplantarum, L. delbrueckii subsp. delbrueckii, L. reuteri, L. murinus, L. agilis, L.delbrueckii subsp. lactis, L. salivarus subsp. salicinus, L. viridenscens, L. amylophilus.

Palavras-chave: Lactobacillus spp., cama de frango, frango.

Lactobacillus species are a bacterial group with morphologic, metabolic and physiologic common characteristics. They are Gram-positive, non-motile, non spore-forming, catalase negative, cytochrome absent, facultative anaerobes, aerotolerant, nutritionally demanding, acid-tolerant, with strictly fermentative metabolism, and lactic acid is the main product of carbohydrates fermentation (6). They are found in nutritionally rich habitats, such as milk, meat, vegetables, grains and in enteric normal microbiota from different animal species (2).

Studies on the intestinal bacteria and pathogen control by intestinal microbiota gained great interest in avian production in the last years (8). Avian enteric microbiota is formed according to the anatomic and physiologic characteristics conditions of the gastrointestinal tract. The microbial population on broiler litter is extremely diversified due to the continuous deposition of birds' excreta, secretions and desquamations. In this microenvironment, strict aerobes or anaerobes, and facultative anaerobes are the outstanding genera (7).

Lactobacillus colonizes avian enteric tract since the first day of life. These bacteria can colonize the small intestine of chicks, being ileum the segment with greater Lactobacillus diversity (5).

Broiler litter can be defined as vegetal material (wood, shaves, rice husk, coffee straw, sugar cane and other byproducts) that contains excreta (feces and urine), skin debris, feathers and feed residues (1). A "new"litter is the one that has not yet received production of birds, while the so-called "re-used"litter is that which has already received production of bird flocks once or more times.

The aim of this survey was to identify Lactobacillus spp. in broiler litter samples from different poultry-rearing regions in Brazil.

One hundred samples of broiler litters from nine poultry-rearing areas in Brazil were used (Table 1). Each poultry-rearing farm sent 300 to 500 g of broiler litter collected in different points of the broiler houses. The samples were fractionated in 20g packages and kept at 4ºC in a sterile recipient.

Isolation of Lactobacillus species was carried out as follows: 5g of each sample broiler litter were added to 50 ml of MRS broth (Difco, Detroit, ML, USA) and kept at 37ºC for 24 hours. Afterwards, the sample was grown on MRS agar and maintained at 37ºC for 24 to 72 hours. Identification of bacteria was done as described by Klander and Weiss (6). The bacteria were lyophilized and kept at -20ºC.

Out of the one hundred samples studied, twenty-three (23%) Lactobacillus species were isolated: L. plantarum (39.1%), L. casei subsp. pseudoplantarum (17.4%), L. delbrueckii subsp. delbrueckii (8.7%), L. reuteri (8.7%), L. agilis (4.3%), L. murinus (4.3%), L. delbrueckii subsp. lactis (4.3%), L. salivarus subsp. salicinius (4.3%), L. viridenscens (4.3%) and L. amylophilus (4.3%).

Several factors may interfere on Lactobacillus viability in the litter, such as litter pH, bird age, environmental temperature, state of re-use and others (3). In this study, the isolation of Lactobacillus species was not affected by the litter original kind, nor by the times the litter was reused (Table 1).

The microbial population in broiler litter is highly diversified. Regarding the bacterial population on the enteric tract, Enterobacteriaceae, Lactobacillus, Micrococcus, Pseudomonas, Bacillus, Sarcinae, Clostridium and anaerobic bacteria are the main bacterial groups (3).

In this study, L. agilis, L. murinus and L. viridiensis were isolated from broiler litters. These species neither are commonly found in avian environments, nor in commercial birds' enteric tract, as some other Lactobacillus species, which are often isolated from avian gut. Therefore, their importance for the enteric microbiota balance, as well as their role in the development of intensively raised birds are being speculated (4).

These results showed the diversity of Lactobacillus species isolated in broiler litter samples obtained from different poultry-rearing areas in Brazil. Other studies should be carried out in order to better understand the importance of these species upon broiler intestinal microbiota regulation, reduction of ammonium in the shed, and their interference on the elimination of pathogens, such as Salmonella and Campylobacter sp.

ACKNOWLEDGEMENTS

We are grateful to FAPESP for the financial support to this study (grants 97/3638 and 99/05087-8).

Submitted: May 05, 2003; Approved: September 01, 2003

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Publication Dates

• Publication in this collection
  04 Aug 2004
• Date of issue
  July 2003

History

• Received
  05 May 2003
• Accepted
  01 Sept 2003