Transferrina como marcador de maturação pulmonar em cordeiros nascidos a termo ou prematuros / Transferrin as a marker of pulmonary maturation in lambs born at term or premature

O objetivo do estudo foi procurar proteínas de fase aguda que possam indicar sinais de maturação no neonato prematuro, por meio da quantificação sérica delas. Identificou-se a imunoglobulina A, a ceruloplasmina, a haptoglobina, a glicoproteína ácida, a transferrina, a albumina e as imunoglobulinas G de cadeias leve e pesada, pela comparação do perfil dos proteinogramas de cordeiros nascidos a termo com os prematuros submetidos a diferentes protocolos terapêuticos, a fim de estimular a atividade respiratória. Constituíram-se seis grupos: PN (n= 9): nascidos de parto normal; CN (n= 7): nascidos de cesariana em tempo normal de gestação; CP (n= 6): nascidos de cesariana prematura sem nenhum tipo de tratamento; DEX (n= 9): prematuros cujas mães receberam dexametasona pré-parto; SURF (n= 6): prematuros tratados com surfactante; e DEXSURF (n= 6): prematuros tratados com surfactante cujas mães receberam dexametasona pré-parto. As avaliações foram realizadas nos momentos imediatamente após o nascimento (M0), após 24 (M24) e após 48 horas (M48). As amostras foram processadas por meio de eletroforese em gel de poliacrilamida contendo dodecil sulfato de sódio (SDS-PAGE). A albumina, as imunoglobulinas e a proteína total dos cordeiros tiveram elevação após a ingestão de colostro. Maiores valores séricos de transferrina são referentes a maior período gestacional, podendo essa proteína ser utilizada como marcador de maturação neonatal.(AU)

The aim of this study was to search for acute phase proteins that could indicate signs of maturation in the premature neonate by quantifying them in serum. Immunoglobulin A, ceruloplasmin, haptoglobin, acid glycoprotein, tranferrin, albumin, light and heavy chain immunoglobulin G were quantified, comparing the profile of proteinograms from term to preterm lambs submitted to different protocols that stimulate respiratory activity. Six groups were used: PN (n= 9): born from normal birth; CN (n= 7): born from caesarean section at normal time of gestation; CP (n= 6): born from premature cesarean without any type of treatment; DEX (n= 9) preterm whose mothers received prepartum dexamethasone; SURF (n= 6) preterm treated with surfactant; DEXSURF (n= 6): preterm treated with surfactant whose mothers received prepartum dexamethasone. The evaluations were performed immediately after birth (M 0), after 24 and 48 hours (M 24 and M 48). Samples were processed with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Albumin, immunoglobulins, and serum total protein of the lambs were elevated, after colostrum ingestion. Higher serum transferrin values refer to a longer gestational period, and this protein may be used as a marker of neonatal maturation.(AU)

Colorectal premalignancy is associated with consensus molecular subtypes 1 and 2.

Background: Gene expression-based profiling of colorectal cancer (CRC) can be used to identify four molecularly homogeneous consensus molecular subtype (CMS) groups with unique biologic features. However, its applicability to colorectal premalignant lesions remains unknown. Patients and methods: We assembled the largest transcriptomic premalignancy dataset by integrating different public and proprietary cohorts of adenomatous and serrated polyps from sporadic (N = 311) and hereditary (N = 78) patient populations and carried out a comprehensive analysis of carcinogenesis pathways using the CMS random forest (RF) classifier. Results: Overall, transcriptomic subtyping of sporadic and hereditary polyps revealed CMS2 and CMS1 subgroups as the predominant molecular subtypes in premalignancy. Pathway enrichment analysis showed that adenomatous polyps from sporadic or hereditary cases (including Lynch syndrome) displayed a CMS2-like phenotype with WNT and MYC activation, whereas hyperplastic and serrated polyps with CMS1-like phenotype harbored prominent immune activation. Rare adenomas with CMS4-like phenotype showed significant enrichment for stromal signatures along with transforming growth factor-ß activation. There was a strong association of CMS1-like polyps with serrated pathology, right-sided anatomic location and BRAF mutations. Conclusions: Based on our observations made in premalignancy, we propose a model of pathway activation associated with CMS classification in colorectal carcinogenesis. Specifically, while adenomatous polyps are largely CMS2, most hyperplastic and serrated polyps are CMS1 and may transition into other CMS groups during evolution into carcinomas. Our findings shed light on the transcriptional landscape of premalignant colonic polyps and may help guide the development of future biomarkers or preventive treatments for CRC.

Surfaceome profiling enables isolation of cancer-specific exosomal cargo in liquid biopsies from pancreatic cancer patients.

Background: Detection of circulating tumor DNA can be limited due to their relative scarcity in circulation, particularly while patients are actively undergoing therapy. Exosomes provide a vehicle through which cancer-specific material can be enriched from the compendium of circulating non-neoplastic tissue-derived nucleic acids. We carried out a comprehensive profiling of the pancreatic ductal adenocarcinoma (PDAC) exosomal 'surfaceome' in order to identify surface proteins that will render liquid biopsies amenable to cancer-derived exosome enrichment for downstream molecular profiling. Patients and methods: Surface exosomal proteins were profiled in 13 human PDAC and 2 non-neoplastic cell lines by liquid chromatography-mass spectrometry. A total of 173 prospectively collected blood samples from 103 PDAC patients underwent exosome isolation. Droplet digital PCR was used on 74 patients (136 total exosome samples) to determine baseline KRAS mutation call rates while patients were on therapy. PDAC-specific exosome capture was then carried out on additional 29 patients (37 samples) using an antibody cocktail directed against selected proteins, followed by droplet digital PCR analysis. Exosomal DNA in a PDAC patient resistant to therapy were profiled using a molecular barcoded, targeted sequencing panel to determine the utility of enriched nucleic acid material for comprehensive molecular analysis. Results: Proteomic analysis of the exosome 'surfaceome' revealed multiple PDAC-specific biomarker candidates: CLDN4, EPCAM, CD151, LGALS3BP, HIST2H2BE, and HIST2H2BF. KRAS mutations in total exosomes were detected in 44.1% of patients undergoing active therapy compared with 73.0% following exosome capture using the selected biomarkers. Enrichment of exosomal cargo was amenable to molecular profiling, elucidating a putative mechanism of resistance to PARP inhibitor therapy in a patient harboring a BRCA2 mutation. Conclusion: Exosomes provide unique opportunities in the context of liquid biopsies for enrichment of tumor-specific material in circulation. We present a comprehensive surfaceome characterization of PDAC exosomes which allows for capture and molecular profiling of tumor-derived DNA.

High prevalence of mutant KRAS in circulating exosome-derived DNA from early-stage pancreatic cancer patients.

Background: Exosomes arise from viable cancer cells and may reflect a different biology than circulating cell-free DNA (cfDNA) shed from dying tissues. We compare exosome-derived DNA (exoDNA) to cfDNA in liquid biopsies of patients with pancreatic ductal adenocarcinoma (PDAC). Patients and methods: Patient samples were obtained between 2003 and 2010, with clinically annotated follow up to 2015. Droplet digital PCR was performed on exoDNA and cfDNA for sensitive detection of KRAS mutants at codons 12/13. A cumulative series of 263 individuals were studied, including a discovery cohort of 142 individuals: 68 PDAC patients of all stages; 20 PDAC patients initially staged with localized disease, with blood drawn after resection for curative intent; and 54 age-matched healthy controls. A validation cohort of 121 individuals (39 cancer patients and 82 healthy controls) was studied to validate KRAS detection rates in early-stage PDAC patients. Primary outcome was circulating KRAS status as detected by droplet digital PCR. Secondary outcomes were disease-free and overall survival. Results: KRAS mutations in exoDNA, were identified in 7.4%, 66.7%, 80%, and 85% of age-matched controls, localized, locally advanced, and metastatic PDAC patients, respectively. Comparatively, mutant KRAS cfDNA was detected in 14.8%, 45.5%, 30.8%, and 57.9% of these individuals. Higher exoKRAS MAFs were associated with decreased disease-free survival in patients with localized disease. In the validation cohort, mutant KRAS exoDNA was detected in 43.6% of early-stage PDAC patients and 20% of healthy controls. Conclusions: Exosomes are a distinct source of tumor DNA that may be complementary to other liquid biopsy DNA sources. A higher percentage of patients with localized PDAC exhibited detectable KRAS mutations in exoDNA than previously reported for cfDNA. A substantial minority of healthy samples demonstrated mutant KRAS in circulation, dictating careful consideration and application of liquid biopsy findings, which may limit its utility as a broad cancer-screening method.

Partial hepatectomy in a calf with an omphalocele: case report / Hepatectomia parcial em bezerra portadora de onfalocele: relato de caso

An omphalocele is a defect in the closure of the abdominal wall during the embryonic stage, preventing the return of some abdominal organs to the cavity. In stillborn animals, the involvement of the liver in the omphalocele has been reported. The aim of the present study is to report the success of a partial liver resection in a female bovine at two days of age which presented an omphalocele at birth. Surgical intervention was indicated to reduce and suture the abdominal wall. During the surgical procedure, the presence of hard, reddish tissue was noted inside the sac covered by the amniotic membrane, with characteristics consistent with hepatic parenchyma. Due to the impossibility of reducing the contents added to the suggestion of tissue infection, we elected to perform a partial resection. The histopathological examination confirmed that the resected tissue was of the hepatic parenchyma. Antibiotic and systemic anti-inflammatory therapies were performed post-surgery. Fifteen days after surgery, infection was detected at the surgical site in addition to abdominal wound dehiscence; surgical reintervention was performed. On physical examination, carried out 4 months after the second operation, the heifer presented normal development for the species. In conclusion, omphalocele may contain liver. Ectopic liver is an extremely rare condition. Surgical treatment in the presented case focused on umbilicus with exploration of the abdominal cavity appeared to be sufficient.(AU)

A onfalocele é um defeito no fechamento da parede abdominal durante a fase embrionária, não permitindo o retorno de alguns órgãos abdominais à cavidade. O envolvimento do fígado em onfalocele tem sido relatado, porém em animais natimortos. Com o presente estudo, objetiva-se relatar uma ressecção parcial do fígado realizada com sucesso em uma fêmea bovina com dois dias de idade apresentando onfalocele ao nascimento. Foi indicada a intervenção cirúrgica para a redução e rafia da parede abdominal. Durante o procedimento cirúrgico, notou-se, no interior do saco recoberto pela membrana amniótica, a presença de tecido firme, de coloração avermelhada, com características compatíveis com parênquima hepático. Devido à impossibilidade de redução do conteúdo e sugestão de infecção do tecido encontrado, optou-se pela ressecção parcial dele. O exame histopatológico confirmou tratar-se de parênquima hepático o tecido resseccionado. No pós-cirúrgico, realizaram-se terapias antibiótica e antiinflamatória sistêmicas. Após 15 dias do procedimento cirúrgico, foi detectada infecção do sítio operatório e deiscência da sutura abdominal, sendo realizada reintervenção cirúrgica. No exame físico, realizado no animal após quatro meses da segunda intervenção cirúrgica, constatou-se desenvolvimento normal para a espécie. Conclui-se que o fígado pode ser conteúdo da onfalolocele, sendo uma condição extremamente rara. O tratamento cirúrgico no caso apresentado, com exploração da cicatriz umbilical e da cavidade abdominal, foi suficiente.(AU)

Minimally invasive genomic and transcriptomic profiling of visceral cancers by next-generation sequencing of circulating exosomes.

BACKGROUND: The ability to perform comprehensive profiling of cancers at high resolution is essential for precision medicine. Liquid biopsies using shed exosomes provide high-quality nucleic acids to obtain molecular characterization, which may be especially useful for visceral cancers that are not amenable to routine biopsies. PATIENTS AND METHODS: We isolated shed exosomes in biofluids from three patients with pancreaticobiliary cancers (two pancreatic, one ampullary). We performed comprehensive profiling of exoDNA and exoRNA by whole genome, exome and transcriptome sequencing using the Illumina HiSeq 2500 sequencer. We assessed the feasibility of calling copy number events, detecting mutational signatures and identifying potentially actionable mutations in exoDNA sequencing data, as well as expressed point mutations and gene fusions in exoRNA sequencing data. RESULTS: Whole-exome sequencing resulted in 95%-99% of the target regions covered at a mean depth of 133-490×. Genome-wide copy number profiles, and high estimates of tumor fractions (ranging from 56% to 82%), suggest robust representation of the tumor DNA within the shed exosomal compartment. Multiple actionable mutations, including alterations in NOTCH1 and BRCA2, were found in patient exoDNA samples. Further, RNA sequencing of shed exosomes identified the presence of expressed fusion genes, representing an avenue for elucidation of tumor neoantigens. CONCLUSIONS: We have demonstrated high-resolution profiling of the genomic and transcriptomic landscapes of visceral cancers. A wide range of cancer-derived biomarkers could be detected within the nucleic acid cargo of shed exosomes, including copy number profiles, point mutations, insertions, deletions, gene fusions and mutational signatures. Liquid biopsies using shed exosomes has the potential to be used as a clinical tool for cancer diagnosis, therapeutic stratification and treatment monitoring, precluding the need for direct tumor sampling.

Clinicopathologic characteristics and gene expression analyses of non-KRAS 12/13, RAS-mutated metastatic colorectal cancer.

BACKGROUND: KRAS mutations in codons 12 and 13 are present in ∼40% of all colorectal cancers (CRC). Activating mutations in codons 61 and 146 of KRAS and in codons 12, 13, and 61 of NRAS also occur but are less frequent. The clinicopathologic features and gene expression profiles of this latter subpopulation of RAS-mutant colorectal tumors have not yet been clearly defined but in general are treated similarly to those with KRAS 12 or 13 mutations. PATIENTS AND METHODS: Records of patients with metastatic CRC (mCRC) treated at MD Anderson Cancer Center between December 2000 and August 2012 were reviewed for RAS (KRAS or NRAS) and BRAF mutation status, clinical characteristics, and survival outcomes. To study further with an independent cohort, data from The Cancer Genome Atlas were analyzed to define a gene expression signature for patients whose tumors feature these atypical RAS mutations and explore differences with KRAS 12/13-mutated colorectal tumors. RESULTS: Among the 484 patients reviewed, KRAS 12/13, KRAS 61/146, NRAS, and BRAF mutations were detected in 47.7%, 3.0%, 4.1%, and 7.4%, respectively, of patients who were tested for each of these aberrations. Lung metastases were more common in both the KRAS 12/13-mutated and atypical RAS-mutated cohorts relative to patients with RAS/BRAF wild-type tumors. Gene expression analyses revealed similar patterns regardless of the site of RAS mutation, and in silico functional algorithms predicted that KRAS and NRAS mutations in codons 12, 13, 61, and 146 alter the protein function and drive tumorgenesis. CONCLUSIONS: Clinicopathologic characteristics, survival outcomes, functional impact, and gene expression profiling were similar between patients with KRAS 12/13 and those with NRAS or KRAS 61/146-mutated mCRC. These clinical and bioinformatic findings support the notion that colorectal tumors driven by these RAS mutations are phenotypically similar.

Aspectos morfométricos da cicatrização do cólon descendente de eqüinos submetidos a enterorrafias aposicionais com poliglactina 910 e com cianoacrilato / Morphometric aspects of the healing of descending colon of horses submitted to appositional suture techniques with polyglactin 910 and with cyanoacrilate

Foi realizado estudo morfométrico do cólon descendente de 15 eqüinos hígidos, nove fêmeas e seis machos, submetidos a enterorrafias aposicionais com fio de poliglactina 910 ou com cianoacrilato. Os animais foram distribuídos aleatoriamente em cinco grupos de três animais cada e sacrificados aos três, sete, 14, 35 e 70 dias de pós-operatório para colheita dos segmentos intestinais operados e posterior processamento histológico de rotina. A análise morfométrica foi feita em microscópio óptico adaptado a um sistema de análise de imagens, onde se quantificou o infiltrado inflamatório, mediante contagens de neutrófilos, eosinófilos, macrófagos, linfócitos e plasmócitos, não sendo observadas diferenças (P>0,05) entre as enterorrafias. Houve diferenças significativas entre os dias de pós-operatório para as contagens de neutrófilos, eosinófilos, macrófagos e linfócitos. Também se mensuraram as espessuras das regiões operadas (cicatriz) e das áreas contíguas não operadas (controle), registrando-se diferenças entre os dias de pós-operatório e as regiões (P<0,05), mas não entre as enterorrafias. O fio de poliglactina 910 e o cianoacrilato apresentam a mesma eficácia para serem usados em enterorrafias aposicionais do cólon descendente de eqüinos.

A morphometric study was carried out in descending colon of 15 healthy horses, nine females and six males, submitted to appositional suture techniques with polyglactin 910 or with cyanoacrilate. The animals were randomly distributed in five groups of three animals each and sacrificed on postoperative days 3, 7, 14, 35 or 70 in order to collect the operated intestinal segments. The morphometric evaluation was done with an optic microscope adapted to an image analysis system, where was counted the inflammatory infiltrated, through neutrophils, eosinophils, macrophages, lymphocytes and plasmacytes counting. No difference (P>0.05) was observed between the two suture techniques. There were significant differences between postoperative days related to the counting of neutrophils, eosinophils, macrophages and lymphocytes. It was also measured the thickness of operated regions (scar) and non operated nearby areas (control); it registered differences between the postoperative days and the regions (P<0.05), but not between the sutures. The polyglactin 910 and the cyanoacrilate showed the same efficacy to be used in appositional enterorrhaphies in equine descending colon.

Estudo comparativo da cicatrizaçäo de enterorrafias em planos aposicional e invaginante no cólon descendente de eqüinos / Comparative study between appositional and inverting pattern for closure of descending colon enterotomies in equines

O estudo foi conduzido com o objetivo de comparar o processo cicatricial do cólon descendente de eqüinos, quando submetido a duas técnicas de sutura, uma em plano aposicional e outra em dois planos com invaginaçäo. Foram utilizados 15 eqüinos sadios e a intervençäo cirúrgica foi realizada com os animais em decúbito lateral direito, sob anestesia geral inalatória. O acesso à cavidade abdominal foi feito pelo flanco esquerdo, e após exteriorizaçäo do cólon descendente, realizaram-se duas enterotomias de 5cm de extensäo cada, distanciadas 20cm uma da outra. Os animais foram aleatoriamente distribuídos em cinco grupos de três animais cada e sacrificados aos 3, 7, 14, 21 e 35 dias pós-operatório. O exame necroscópico revelou a presença de aderências nos dois padröes de sutura. Em nove eqüinos foram observadas aderências no padräo de sutura aposicional (60,0 por cento +/- 12,6) e em quatro no padräo com invaginaçäo (26,6 por cento +/- 11,4). Näo foi verificada interferência com o trânsito intestinal em nenhum paciente. O exame histopatológico revelou regeneraçäo da camada mucosa a partir do sétimo dia de pós-operatório em ambos os tipos de satura. O fio de poliglactina 910 empregado nas enterorrafias do cólon descendente se mostrou resistente, de fácil manuseio e com boa capacidade para manter os nós aplicados, causando moderada reaçäo inflamatória do tipo granulomatosa

Avaliaçäo da eficiência de um probiótico no controle de diarréia e no ganho de peso de bezerros / Evaluation of the efficiency of a probiotic in the control of diarrhea and weight gain in calves

Foram utilizadas 99 vacas prenhes distribuídas em oito grupos que receberam os seguintes tratamentos: grupo I, com 29 vacas näo vacinadas e seus bezerros que näo receberam probiótico, ficando como controle; grupo II, com 10 vacas vacinadas e seus bezerros que näo receberam probiótico; grupos III,IV e V, com 10 animais cada, vacas vacinadas e seus bezerros que receberam probiótico durante 5, 15 e 30 dias, respectivamente; os grupos VI, VII e VIII, com 10 animais cada, vacas näo vacinadas e seus bezerros que receberam probiótico durante 5, 15 e 30 dias, respectivamente. Cada animal dos grupos vacinados recebeu duas doses vacinais contendo os pili K99 e A14 de Escherichia coli na dose de 5,0ml por via subcutânea. O probiótico contendo Ruminobacter amylophilum, Ruminobacter succinogenes, Succinovibrio dextrinosolvens, Bacillus cereus, Lactobacillus acidophilus e Streptococcus faecium, na dose de 3,0X10 elevado a oitava potência células vivas (UFC) de cada amostra em 250ml de leite, era administrado por via oral. Os animais foram observados diariamente e foram determinados os títulos de anticorpos anti-K99 e anti-A14 no soro e no colostro. Anotaram-se os pesos dos bezerros ao nascimento e aos 30 dias. Os resultados mostraram que a associaçäo de vacina com probiótico administrado por 15 e 30 dias foram os tratamentos mais eficientes no controle da diarréia e ganho de peso

Constituintes sangüíneos de bovinos recém-nascidos das raça Nelore (Bos indicus) e Holandesa (Bos taurus) e de bubalinos (Bubalus bubalis) da raça Murrah / Blood constituents of the newborn Nelore cattle (Bos indicus), Holstein cattle (Bos taurus), and Murrah buffalo (Bubalus bubalis)

Foram examinadas amostras de sangue de bovinos recém-nascidos das raças Nelore e Holandesa e de bubalinos da raça Murrah, obtidas no dia do nascimento e aos 15, 30 e 45 dias de idade. Os resultados indicaram que: 1. a contagem de hemácias de bubalinos foi inferior à de bovinos nos primeiros 30 dias de idade, no entanto o teor de hemoglobina de búfalos foi superior à de bovinos; 2. as contagens de leucócitos, neutrófilos e linfócitos foi maior em bubalinos que em bovinos, e decresceram com o avanço da idade, diferentemente do número de eosinófilos e de monócitos que manifestaram tendência de elevaçäo no período; 3. os maiores valores de fibrinogênio foram constatados no dia do nascimento, enquanto os níveis das proteínas totais séricas tenderam à elevaçäo em funçäo da idade; 4. os níveis glicêmicos diminuíram com o avanço da idade e a glicemia de bubalinos foi inferior à de bovinos Nelore; 5. as concentraçöes de ferro, uréia e aspartato aminotransferase de bubalinos superaram as de bovinos, com maiores valores no dia do nascimento, e perfil semelhante foi notado com a creatinina; 6. os níveis de bilirrubina e alanina aminotransferase de bovinos foram superiores aos de búfalos, com maior valor ao nascimento; 7. a partir de 15 dias de idade, os bovinos apresentaram atividade de fosfatase alcalina superior à de bubalinos e em todos os grupos ocorreu queda da atividade enzimática ao longo do tempo; 8. a atividade de gamaglutamiltransferase foi maior no dia do nascimento com acentuado decréscimo com o avanço da idade; 9. os níveis séricos de cálcio e fósforo apresentaram oscilaçöes de pouca intensidade, enquanto a concentraçäo de magnésio foi superior em búfalos, com menores valores ao nascimento

Constituintes sangüíneos de bovinos lactantes, desmamados e adultos da raça Nelore (Bos indicus) e Holandesa (Bos taurus) e de bubalinos (Bubalus bubalis) da raça Murrah / Blood constituents of lactating, weaned and adult Nelore cattle (Bos indicus), and Holstein cattle (Bos taurus), and Murrah buffalo (Bubalus bubalis)

Foram analisadas amostra de sangue de bovinos lactentes, desmamados e adultos das raças Nelore e Holandesa e de bubalinos da raça Murrah. As contagens de hemácias e de linfócitos, o pH sagüíneos, os teores de fibrinogênio plasmático e os níveis séricos de cálcio e fósforo foram similares em bovinos e bubalinos. As concentraçöes de proteínas totais, uréia, creatinina, hemoglobina, magnésio e ferro e as atividades da enzima aspartato aminotransferase de bubalinos foram superiores às de bovinos. As contagens de leucócitos de bovinos desmamados da raça Holandesa foram superiores às da raça Nelore e semelhantes às dos bubalinos. O teor de bilirrubina de bubalinos foi menor que o de bovinos. A glicemia de animais lactentes e adultos näo foi influenciada por raça ou sexo. A atividade de fosfatase alcalina foi inferior em bubalinos desmamados. A gamaglutamiltransferase apresentou atividades superiores em búfalos lactentes e desmamados

A comparative study of the efficiency of a pro-biotic and the anti-K99 and anti-A14 vaccines in the control of diarrhea in calves in Brazil.

A total of 99 pregnant cows were divided into eight groups submitted to the following treatments: group I (n = 29) consisted of unvaccinated cows whose calves did not receive a probiotic and was used as control. Group II (n = 10) consisted of vaccinated cows whose calves did not receive a probiotic. Groups III, IV and V (n = 10 neach) consisted of vaccinated cows whose calves received a probiotic for 5, 15 and 30 days, respectively. Groups VI, VII and VIII (n = 10 each) consisted of unvaccinated cows whose calves received a probiotic for 5, 15 and 30 days, respectively. Each animal in the vaccinated groups received two 5.0 ml vaccine doses containing pili K99 and A14 of Escherichia coli by the subcutaneous route. The probiotic containing Lactobacillus acidophilus at the dose of 2.0 x 10(8) live cells in 250 ml milk, was administered orally. All animals were observed clinically and bacteriologically and anti-K99 and anti-A14 antibody titers were determined in serum and colostrum. Mean calf weight was measured at birth and at 30 days of age. The results showed that a combination of the vaccine with the probiotic administered for 15 and 30 days was the most efficient treatment for the control of diarrhea.