Protease-activated receptor 2, dipeptidyl peptidase I, and proteases mediate Clostridium difficile toxin A enteritis

BACKGROUND & AIMS: We studied the role of protease-activated receptor 2 (PAR(2)) and its activating enzymes, trypsins and tryptase, in Clostridium difficile toxin A (TxA)-induced enteritis. METHODS: We injected TxA into ileal loops in PAR(2) or dipeptidyl peptidase I (DPPI) knockout mice or in wild-type mice pretreated with tryptase inhibitors (FUT-175 or MPI-0442352) or soybean trypsin inhibitor. We examined the effect of TxA on expression and activity of PAR(2) and trypsin IV messenger RNA in the ileum and cultured colonocytes. We injected activating peptide (AP), trypsins, tryptase, and p23 in wild-type mice, some pretreated with the neurokinin 1 receptor antagonist SR140333. RESULTS: TxA increased fluid secretion, myeloperoxidase activity in fluid and tissue, and histologic damage. PAR(2) deletion decreased TxA-induced ileitis, reduced luminal fluid secretion by 20%, decreased tissue and fluid myeloperoxidase by 50%, and diminished epithelial damage, edema, and neutrophil infiltration. DPPI deletion reduced secretion by 20% and fluid myeloperoxidase by 55%. In wild-type mice, FUT-175 or MPI-0442352 inhibited secretion by 24%-28% and tissue and fluid myeloperoxidase by 31%-71%. Soybean trypsin inhibitor reduced secretion to background levels and tissue myeloperoxidase by up to 50%. TxA increased expression of PAR(2) and trypsin IV in enterocytes and colonocytes and caused a 2-fold increase in Ca(2+) responses to PAR(2) AP. AP, tryptase, and trypsin isozymes (trypsin I/II, trypsin IV, p23) caused ileitis. SR140333 prevented AP-induced ileitis. CONCLUSIONS: PAR(2) and its activators are proinflammatory in TxA-induced enteritis. TxA stimulates existing PAR(2) and up-regulates PAR(2) and activating proteases, and PAR(2) causes inflammation by neurogenic mechanisms.

Sources and spread of thermophilic Campylobacter spp. during partial depopulation of broiler chicken flocks

The practice of partial depopulation or ‘thinning’, i.e. early removal of a proportion of birds from a commercial broiler flock, is a reported risk factor for Campylobacter colonization of residual birds because of the difficulty in maintaining biosecurity during the process. Therefore, the effect of this practice was studied in detail for 51 target flocks, each at a different growing farm belonging to one of seven major poultry companies throughout the United Kingdom. On 21 of these farms, the target flock was already colonized by Campylobacter and at slaughter all cecal samples examined were positive, with a mean of log10 8 cfu / g. A further 27 flocks became positive within 2 – 6 days of the start of thinning and had similarly high levels of cecal carriage at slaughter. Just prior to the thinning process, Campylobacter could be isolated frequently from the farm driveways, transport vehicles, equipment and personnel. Strains from seven such farms on which flocks became colonized after thinning were examined by PFGE typing. The study demonstrated an association between strains occurring at specific sampling sites and those isolated subsequently from the thinned flocks. There were also indications that particular strains had spread from one farm to another, when the farms were jointly company-owned and served by the same bird-catching teams and / or vehicles. The results highlighted the need for better hygiene control in relation to catching equipment and personnel, and more effective cleaning and disinfection of vehicles, and bird-transport crates.

Survey of chicken abattoir for the presence of Campylobacter jejuni and Campylobacter coli

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Incidence of Campylobacter in pigs with and without diarrhea

Um total de 200 suínos (1 - 21 semanas de idade), originários de cinco criações localizadas no Estado de São Paulo, Brasil foi dividido em dois grupos de 100 animais caracterizando-se o grupo G1 de animais com diarréia e G2, sem diarréia. Campylobacter foi isolado em 43% das amostras provenientes de G1 e 34% de G2. O microrganismo foi mais frequentemente encontrado em leitões na faixa de 0 a 4 semanas de idade. Campylobacter coli foi a espécie mais comumente observada em G1 (44,2%) e em G2 (32,4%), seguido por Campylobacter jejuni/ coli com 16,3% em G1 e 23,5% em G2. As contagens de Campylobacter foram significativamente maiores (p < 0,01) em G1 ( £ 108 UFC/g) do que em G2 (£ 104 UFC/g), fato que sugere que o microrganismo pode, pelo menos, atuar no agravamento do processo diarréico.

Detection of turkey astrovirus in young poults affected with poult enteritis complex in Brazil

Reverse transcription polymerase chain reaction (RT-PCR) of turkey astrovirus (TAstV) capsid and polymerase genes was applied to the bursa of Fabricius (BF), thymus (TH), spleen (SP) and cloacal swabs (CS) of young poults with "Poult enteritis complex" (PEC). The histological lesions included atrophy, lymphoid depletion, cellular infiltration and necrosis of the BF, TH and SP, respectively. The RT-PCR reactions were positive for the polymerase gene of TAstV-2 in all 100 CSs, 7 out of 10 of BFs and 10 out of 20 THs and SPs, respectively. Five out of 10 THs and SPs samples, considered to be negative by RT-PCR, were positive when specific primers designed for the TAstV-2 capsid gene were applied. This is the first description of turkey astrovirus infection presenting PEC in Latin America.

Occurrence of Campylobacter in the genitals of teaser bulls maintained at an embryo transfer center

Em central de transferência de embriões, após os procedimentos de reconhecimento do cio em 37 vacas receptoras, através de quatro rufiões vasectomizados, observou-se que 83% delas apresentavam retorno ao cio e algum corrimento serofibrinoso. Nos exames bacteriológicos realizados nos lavados prepuciais dos rufiões foi isolado o Campylobacter fetus subsp. venerealis em todos, fato que, analisado associadamente com o retorno ao cio das vacas receptoras, é indicativo da ocorrência de campilobacteriose no plantel. Essa ocorrência demonstra a necessidade de medidas eficazes de planejamento de saúde animal, pela utilização de rufiões com desvio lateral do pênis. Uma vez impossibilitado o contato sexual, seria impedida a transmissão do agente durante o coito. Torna-se imperioso consignar que a prática da prevenção racional de enfermidades continua sendo o procedimento mais econômico para uma produtividade animal mais rentável.

The comparison of the Butzler medium, filtration technique and their association with isolation of Campylobacter ssp

Do total de 622 amostras de fezes, sendo 200 de suínos, 220 de bezerros e 202 de cães, Campylobacter foi isolado em 178 amostras. Destas, o agente foi identificado em 64 (36%) amostras, isoladas somente em meio seletivo de Butzler (MSB); em 34 (19%) o microorganismo foi isolado a partir da técnica de filtração (TF) e em 80 (45%), através de ambos os procedimentos (MSB+TF). A comparação entre as proporções de positividade, pelas diferentes técnicas, revelou significância (c2 = 9.184; p > 0,001) sendo a MSB (36%) a mais eficiente comparada com a TF (19%) O uso associado de ambos os procedimentos proporcionou a positividade mais elevada de isolamento com 45%.

ENTEROTOXIN PRODUCTION IN STRAINS OF CAMPYLOBACTER-JEJUNI, ISOLATED FROM SWINE DIARRHEA

One hundred and twenty-six strains of Campylobacter jejuni isolated from swine with diarrhea were examined for the production of enteroxin, by culture filtrate inoculation into ileal loops in several animal species.Four (3,1 %) of the strains tested produced fluid in ligated ileal loop of albino rabbit, 19 (15 %) in rat and 14(11, 1 %) in swine. By the test of intragastric inoculation in suckling mouse, none of the strains revealed capacity to produce enterotoxin, although two strains were considered suspicious. Twelve (54,5 %) of the strains that induced ileal fluid in rat and swine produced cytotoxic effect in monkey kidney cells (Vero), affecting up to 60 % of the monolayer. However, no alteration was observed in hamster kidney cells (BHK).

Isolation of Campylobacter jejuni from viscera and bile secretion of broiler chickens with diarrhea

Since chickens are important reservoirs of Campylobacter jejuni and their meat is the most frequent route of transmission for human campylobacteriosis, the purpouse of the present study was to investigate the presence of Campylobacter in viscera of chickens with diarrhea, evaluating the frequency of isolation of this microorganism from organs considered to be preferential for its isolation. A total of 107 visceral samples from chickens with diarrhea from different farms in the Ribeirao Preto region, SP, were examined for the presence of Campylobacter jejuni. The material consisted of 73 livers and 34 spleens, plus 29 bile secretion samples. The frequency of Campylobacter jejuni isolation was 54.79% for the liver samples, 35,29% for the spleens and 6,89% for the bile secretion samples. The data suggest that, under the conditions of the present study, the liver may be the organ of choice for the isolation of Campylobacter in the presence of diarrhea and liver involvement in chickens.

CAMPYLOBACTER EM CAES COM E SEM DIARREIA: INCIDENCIA E SUSCETIBILIDADE A 21 ANTIMICROBIANOS

Two hundred and four stools of 102 diarrheic (0-12 months of age) and 102 non-diarrheic dogs coming from kennel and ambulatory, respectively, were assayed for the presence of Campylobacter. From the diarrhetic group, 46% of the samples yelded positive Campylobacter isolation mainly found in young animals until 5 months of age (72%), whereas those with age ranging from 6-12 months showed either a lower frequence of the organisms (28%) and a trend of the younger animals to be more charged than the older. 47 Campylobacter strains isolated from the diarrheic group were: C. jejuni biotype 1 (49%); C. jejuni biotyp 2 (11%); C. jejuni/coli (19%); C. coli (8,5%); Campylobacter NARTC group (8,5%) and C. sputoruns (4%). In the non-diarrhetic group, 27 (28%) Campylobacter strains were classified as: C. jejuni biotype 1 (34%) and biotype 2 (28%); C. jejuni/coli (24%) and C. coli (14%). According the biochemical tests, the 1% glycine tolerance test was not taking in account for the differentiation of C. jejuni because 45% of the strains failed in showing characteristic and 3 strains did not reduce the sodium selenite. The biochemical studies also showed phenotipical cross reactions between two Campylobacter NARTC-group strains with the C. jejuni strains, as well as two thermophilic species grew also at 25°C. All the 76 isolates were sensitive to gentamicin, nitrofurantoin and neomycin and resistant to oxacillin and penicillin. Furthermore, for the remaining 16 drugs the populational resistance ranged from 8% to 73% of strains. The presence of Campylobacter in dogs as well their close contact which man makes possible the occurrence of infections as also confirm the campylobacteriosis as an important zoonosis.

Invasive enteritis by Strongyloides stercoralis presenting as acute abdominal distress under corticosteroid therapy.

Overwhelming helminthiasis is still a problem in endemic areas, especially in immunocompromised patients. We report a case of invasive intestinal strongyloidiasis that was clinically expressed as acute abdominal distress in a 73-year-old man from São Paulo who had been receiving methylprednisone, 20 mg/day, for one year for osteoarthritis. A surgical specimen from the ileum revealed invasive enteritis with severe infestation by Strongyloides stercoralis. The patient died of sepsis 6 days after surgery. The possibility of invasive strongyloidiasis should be considered in the differential diagnosis of acute abdominal distress in patients undergoing immunosuppressive therapy.