Chronic lead intoxication in a jaguar (Panthera onca) shot with round lead pellets: case report / Intoxicação crônica por chumbo em uma onça pintada (Panthera onca) alvejada com projéteis de chumbo: relato de caso

Lead is a heavy metal and an important cause of acute or chronic toxicosis in humans, domestic, and wild animals. This report aims to describe a case of chronic lead poisoning in a jaguar (Panthera onca) kept under human care that was rescued from the wild environment. The animal was rescued in poor condition in 2004 and kept under human care at the Belo Horizonte Zoological Garden (Minas Gerais, Brazil) until 2020, when it presented with anorexia, vomiting and ataxia. Over the past years the animal had episodes of anemia and increased serum urea and creatinine. Radiography demonstrated 21 radiopaque projectiles on the right side of the face. At necropsy there were multiple projectiles surrounded by fibrous tissue in the subcutaneous of the right side of the face, fibrinous peritonitis, multiple gastric ulcers, and melena. The lead dosage was performed using the atomic absorption spectrometry technique using renal tissue collected at necropsy, with a result of 908 ppb (µg/kg). The findings of projectiles associated with the dosage of lead above the reference limits allow the diagnosis of chronic intoxication in this case.

Chumbo é um metal pesado e uma causa importante de toxicose crônica no homem e em animais domésticos e selvagens. Este relato teve como objetivo descrever um caso de intoxicação crônica por chumbo em uma onça-pintada (Panthera onca), mantida sob cuidados humanos, que foi resgatada de seu ambiente natural em 2004, em pobre condição corporal. O animal foi encaminhado ao zoológico de Belo Horizonte (MG, Brasil), onde permaneceu até 2020, quando desenvolveu anorexia, vômitos e ataxia. Durante os últimos anos, o animal teve episódios de anemia e aumento da concentração sérica de ureia e creatinina. O exame radiográfico demonstrou 21 projéteis radiodensos na face direita. À necropsia, foram observados vários projéteis envoltos por tecido conjuntivo no subcutâneo da face esquerda, peritonite fibrinosa, múltiplas úlceras gástricas e melena. A concentração de chumbo foi determinada por espectrometria de absorção atômica em amostras de tecido renal, indicando 908 ppb (µg/kg). O achado de projéteis de chumbo associado à elevada concentração de chumbo é compatível com intoxicação crônica por chumbo neste caso.

Evaluation of potential ecological risk assessment of toxic metal (lead) in contaminated meadows in the vicinity of suburban city: soil vs forages vs livestock / Avaliação de risco ecológico em potencial avaliação de metal tóxico (chumbo) em prados contaminados nas proximidades da cidade suburbana: solo x forragens x gado

Heavy metal toxicity is becoming an increasing concern for environmental, human and animal health. The current research analyzed the lead (Pb) contamination in the food chain under three different irrigation sources (ground, canal, and wastewater). Soil, plant and animal samples were collected from the Jhang district of Pakistan and processed with an atomic absorption spectrophotometer. Lead concentration varied in the samples as: 5.22-10.73 mg/kg in soil, 2.46-10.34 mg/kg in forages and 0.736-2.45 mg/kg in animal samples. The observed lead concentration in forage and animal blood samples was higher than the standard limits. The pollution load index (0.640-1.32) in soil showed that lead contamination mainly took place at the wastewater irrigating sites. Bio-concentration factor values (0.313-1.15) were lower than one in all samples except Zea mays, showing that lead metal was actively taken up by Zea mays tissues from the soil. Enrichment factor values ranged from 0.849-3.12, showing a moderate level of lead enrichment. Daily intake and health risk index varied between 0.004-0.020 mg/kg/day and 0.906-4.99, respectively. All the samples showed maximum lead concentration at the wastewater irrigating site compared to the ground or canal water application sites. These results recommended that consistent application of wastewater for forage irrigation must be avoided to prevent health hazards associated with lead in the animal and human food chain. Government must implement adequate strategies to protect the animal and human health from the harms of toxic heavy metals.

A toxicidade de metais pesados está se tornando uma preocupação crescente para a saúde ambiental, humana e animal. A pesquisa atual analisou a contaminação por chumbo (Pb) na cadeia alimentar sob três diferentes fontes de irrigação (solo, canal e águas residuais). Amostras de solo, plantas e animais foram coletadas no distrito de Jhang, no Paquistão, e processadas com um espectrofotômetro de absorção atômica. A concentração de chumbo nas amostras variou em: 5,22-10,73 mg/kg no solo, 2,46-10,34 mg/kg nas forragens e 0,736-2,45 mg/kg nas amostras de animais. A concentração de chumbo observada nas amostras de forragem e sangue animal foi superior aos limites padrão. O índice de carga de poluição (0,640-1,32) no solo mostrou que a contaminação por chumbo ocorreu principalmente em locais de irrigação de águas residuais. Os valores do fator de bioconcentração (0,313-1,15) foram menores que um em todas as amostras, exceto Zea mays, mostrando que o chumbo metálico foi ativamente absorvido pelos tecidos de Zea mays do solo. Os valores do fator de enriquecimento variaram de 0,849-3,12, mostrando um nível moderado de enriquecimento de chumbo. A ingestão diária e o índice de risco à saúde variaram entre 0,004-0,020 mg/kg/dia e 0,906-4,99, respectivamente. Todas as amostras mostraram concentração máxima de chumbo no local de irrigação de águas residuais em comparação com os locais de aplicação de água no solo ou no canal. Esses resultados recomendam que a aplicação consistente de águas residuais para irrigação de forragem deve ser evitada para evitar riscos à saúde associados ao chumbo na cadeia alimentar animal e humana. O governo deve implementar estratégias adequadas para proteger a saúde animal e humana dos danos dos metais pesados tóxicos.

Aborto em bovinos e bubalinos: causas específicas e inespecíficas / Abortion in cattle and buffaloes: specific and non-specific causes

O aborto é caracterizado como a expulsão do feto sem viabilidade para fora do ambiente uterino da progenitora durante o período que vai do 42° dia a o 260° dia. São descritos diversos fatores infecciosos e não infecciosos como etiologia para o aborto. Dentre os infecciosos, destacam-se as infecções por bactérias, vírus, protozoários e fungos. Enquanto as causas não infecciosas englobam falhas de manejo nutricional, intoxicações e medicamentos que ocasionalmente podem levar à morte fetal. Essa mortalidade fetal é causa importante de perdas reprodutivas na cadeia produtiva de animais domésticos, especialmente bovinos e bubalinos. Ambas as espécies participam da produtividade econômica no Brasil e compartilham de diversas doenças que são causas de aborto. Nesse contexto, o objetivo desta revisão foi reunir e discorrer sobre as principais informações concernentes às possíveis etiologias do aborto, sejam elas de origem infecciosa ou não infecciosas nas espécies bovinas e bubalinas no Brasil.

Abortion is characterized as the expulsion of the non-viable fetus outside the mother's uterine environment during the period between the 42nd day to the 260th day. Several infectious and non-infectious factors are described as etiology for miscarriage. Among the infectious, infections by bacteria, viruses, protozoa, and fungi stand out, while non-infectious causes include failures in nutritional management, poisoning, and medications that occasionally can lead to fetal death. This fetal mortality is an important cause of reproductive losses in the production chain of domestic animals, especially cattle and buffaloes. Both species participate in economic productivity in Brazil and share several diseases that are causes of abortion. In this context, this review aimed to gather and discuss the main information concerning the possible etiologies of abortion, whether of infectious or non-infectious origin in bovine and buffalo species in Brazil.

Electrocardiographic, echocardiographic and heart biomarker parameters in sheep experimentally poisoned by Palicourea marcgravii (Rubiaceae) / Parâmetros eletrocardiográficos, ecocardiográficos e de biomarcadores cardíacos em ovinos intoxicados experimentalmente por Palicourea marcgravii (Rubiaceae)

The present study aimed to identify and describe cardiac alterations in sheep experimentally poisoned with Palicourea marcgravii through analysis of serum cardiac biomarkers (serum troponin I and creatine kinase - CK-MB) and electro and echocardiographic assessments to contribute to a better understanding of the poisoning pathophysiology. P. marcgravii is the main plant within a group of 22 species that cause sudden death in Brazil; its toxic principle is sodium monofluoroacetate. Eight healthy crossbreed male sheep, aged between five and twelve months, weighing 14 to 27kg, were evaluated. The animals received 1g kg-1 of P. marcgravii plants orally. The sheep were evaluated before administering the plant (T0) through electro and echocardiography and blood collection to assess cardiac biomarkers (CK-MB and cTnI). Collections and analyses were repeated every four hours until the animal's death. During the study, there was the presence of extravasation of serum troponin I carried out in a qualitative test, with positive values at time T4, and the serum CK-MB biomarker had a peak at T4 and slightly decreased at T8. The electro and echocardiographic examinations showed that the cause of death in these animals was due to acute heart failure characterized by arrhythmias, tachycardia/ventricular fibrillation, drop in cardiac output, left ventricular (LV) systolic dysfunction by the progressive decrease in the LV ejection fraction (EF), decrease in LV fractional shortening (FS), and decrease in aortic flow velocity and aortic flow gradient. This study seems to be the first to evaluate cardiac alterations in sheep poisoned by P. marcgravii through cardiac biomarkers and electro and echocardiographic exams.

O presente estudo objetivou identificar e descrever as alterações cardíacas de ovinos intoxicados experimentalmente por Palicourea marcgravii através das análises de biomarcadores cardíacos séricos (troponina I sérica e a creatinoquinase - MB) e das avaliações eletro e ecocardiográficas contribuindo para o melhor entendimento da fisiopatologia da intoxicação. Palicourea marcgravii é a principal planta dentro de um grupo de 22 espécies que causam "morte súbita" no Brasil e seu princípio tóxico é o monofluoracetato de sódio. Foram utilizados oito ovinos saudáveis machos, sem raça definida, com idades entre cinco e doze meses e peso de 14 a 27kg. Os animais receberam 1g/kg de Palicourea marcgravii por via oral. Os ovinos foram avaliados momentos antes da administração da planta (T0) através de eletro e ecocardiograma e coleta de sangue para avaliação dos biomarcadores cardíacos (CK-MB e cTnI). As análises e coletas foram repetidas a cada quatro horas até o óbito do animal. Durante o estudo observou-se extravasamento de troponina I sérica realizada em teste qualitativo, com valores positivos já em T4, assim como o biomarcador CK-MB sérico teve seu pico de aumento em T4 e em T8 houve uma leve redução. Aos exames eletro e ecocardiográfico foi possível determinar que a causa do óbito nestes animais ocorreu devido à insuficiência cardíaca aguda caracterizada por arritmias, taquicardia/fibrilação ventricular, queda no débito cardíaco, disfunção sistólica do ventrículo esquerdo (VE) pela diminuição progressiva da fração de ejeção (EF) do VE, diminuição na fração de encurtamento (FS) do VE, diminuição da velocidade do fluxo aórtico e do gradiente do fluxo aórtico. Este é o primeiro estudo que avalia as alterações cardiológicas de ovinos intoxicados por P. marcgravii através de biomarcadores cardíacos e exames eletro e ecocardiográficos.

Impact of recreational fishing on seagull (Larus dominicanus) rescued on the coast of the São Paulo state: case report / Impacto da pesca recreativa em gaivota (Larus dominicanus) resgatada no litoral do estado de São Paulo: relato de caso

The seagull (Larus dominicanus) commonly can be observed near human fishing activities, as this species feeds on discards from fishing. This common interaction between a seabird and human fishing activity (both commercial and recreational), provides an easy source of food, but is not without risk for the birds. We report here clinical, radiographic and anatomopathological findings of an esophageal perforation caused by fishing gear in a specimen of Larus dominicanus found alive on a beach in the state of São Paulo which illustrated a harmful effect of stemming from recreational fishing. Clinical examination revealed a nylon thread in the oral cavity with the presence of caseous suggestive of ingestion of lost fishing gear, paralysis of the pelvic limbs and a decrease in the pain reflex associated with sternal decubitus, suggestive of food intoxication. Necroscopic examination revealed two hooks, one in the caudal portion of the esophagus with esophageal and pulmonary perforation, and the other in the muscular stomach. These injuries would have been expected to lead to the death of the bird. These anatomopathological findings confirmed the seriousness of the injuries. This reinforces the importance of studies focused on the interaction of recreational fishing artifacts (hooks, lines) with coastal birds on the Brazilian coast, in order to develop a preventative strategy.(AU)

A gaivota (Larus dominicanus) está intimamente relacionada às atividades pesqueiras, uma vez que possui hábitos generalistas e se alimenta dos descartes provenientes da pesca. Este processo de interação entre aves marinhas e a pesca apre-senta efeitos positivos e negativos, porém, suas relações ecológicas ainda não são bem compreendidas. No presente trabalho, objetivou-se relatar achados clínicos, radiográficos e anatomopatológicos de uma perfuração esofágica ocasionada por petrecho de pesca em um espécime de Larus dominicanus no litoral do estado de São Paulo e levantar a problemática da pesca recrea-tiva na região. Ao exame clínico, observou-se presença de fio de náilon em cavidade oral com presença de cáseo sugestivo de ingestão de petrecho de pesca, paresia de membros pélvicos e diminuição de reflexo doloroso associado a decúbito esternal, sugerindo um quadro de intoxicação alimentar. O exame necroscópico revelou a presença de dois anzóis, um em porção cau-dal do esôfago com perfuração esofágica e pulmonar, e outro em ventrículo. Os achados anatomopatológicos comprovaram a gravidade das lesões causadas pela interação com a pesca recreativa, o que reforça a importância de estudos referentes à intera-ção de artefatos pesqueiros (anzóis, linhas) com aves costeiras do litoral brasileiro, atualmente escassos na literatura científica.(AU)

Plant poisoning containing hydrocyanic acid in cattle in Southern Brazil

Background: Cyanogenic plants accumulate cyanogenic glycosides and release hydrocyanic acid (HCN). In Brazil, therehave been reports of several plants that cause HCN poisoning in animals and lead to a fast death with few clinical signs andlesions on post mortem examination. Some cultivars of Cynodon spp. grasses cause HCN poisoning in cattle in Brazil. Theobjectives of this work were to report the occurrence of deaths by HCN poisoning in cattle as diagnosed by the veterinarypathology laboratory, describe the quantity of HCN in some cultivars of Cynodon spp., as well as, to describe one cultivarof genus Cynodon never reported as poisonous.Materials, Methods & Results: The archives of the Veterinary Pathology Laboratory (LPV) at the Concórdia Campus ofthe Instituto Federal Catarinense (IFC) were reviewed, seeking cases with a diagnosis of hydrocyanic acid poisoning incattle after post mortem examination. The amount of HCN present in some cultivars of the Cynodon genus was quantified due to the high frequency of poisoning cases. From the 1,235 post mortem examinations of cattle 28 (2.27%) werediagnosed with spontaneous hydrocyanic acid poisoning, 17 cases (60.7%) due to ingestion of Prunus sp. or Manihotsp., and 11 cases (39.3%) of Cynodon dactylon ingestion. Most animals were found dead, normally having presented noclinical signs. Macroscopic evaluation mainly showed a severe amount of unchewed and undigested leaves or grass mixedin the ruminal content presenting a bitter almond odor. It was possible to infer that, among cultivars of the Cynodon genus, Florakirk showed the highest levels of HCN compared (P < 0.05) with Star of Puerto Rico, Tifton 68, Tifton 44, andCoast-Cross. Furthermore, Tifton 85 and Jiggs showed undetected levels of HCN...

Plant poisoning containing hydrocyanic acid in cattle in Southern Brazil

Background: Cyanogenic plants accumulate cyanogenic glycosides and release hydrocyanic acid (HCN). In Brazil, therehave been reports of several plants that cause HCN poisoning in animals and lead to a fast death with few clinical signs andlesions on post mortem examination. Some cultivars of Cynodon spp. grasses cause HCN poisoning in cattle in Brazil. Theobjectives of this work were to report the occurrence of deaths by HCN poisoning in cattle as diagnosed by the veterinarypathology laboratory, describe the quantity of HCN in some cultivars of Cynodon spp., as well as, to describe one cultivarof genus Cynodon never reported as poisonous.Materials, Methods & Results: The archives of the Veterinary Pathology Laboratory (LPV) at the Concórdia Campus ofthe Instituto Federal Catarinense (IFC) were reviewed, seeking cases with a diagnosis of hydrocyanic acid poisoning incattle after post mortem examination. The amount of HCN present in some cultivars of the Cynodon genus was quantified due to the high frequency of poisoning cases. From the 1,235 post mortem examinations of cattle 28 (2.27%) werediagnosed with spontaneous hydrocyanic acid poisoning, 17 cases (60.7%) due to ingestion of Prunus sp. or Manihotsp., and 11 cases (39.3%) of Cynodon dactylon ingestion. Most animals were found dead, normally having presented noclinical signs. Macroscopic evaluation mainly showed a severe amount of unchewed and undigested leaves or grass mixedin the ruminal content presenting a bitter almond odor. It was possible to infer that, among cultivars of the Cynodon genus, Florakirk showed the highest levels of HCN compared (P < 0.05) with Star of Puerto Rico, Tifton 68, Tifton 44, andCoast-Cross. Furthermore, Tifton 85 and Jiggs showed undetected levels of HCN...(AU)

Intoxicação por lasalocida em bezerros búfalos no Estado do Pará, Brasil / Lasalocid poisoning in buffalo calves in the state of Pará, Brazil

Background: Ionophore antibiotics are food additives with coccidiostatic or antimicrobial action; they are also used as growth promoters, ruminal pH regulators, volatile fatty acid molar modifiers, and methanogenesis reducers. However, these compounds have the potential to cause microbial resistance, in addition to the risk of intoxication. Ionophore poisoning may be caused by excessive intake, sensitivity of certain animal species, and concomitant use with other drugs. In Brazil, cases of ionophore poisoning in buffalos are rare. This study aims to describe the epidemiological, clinical, and pathological findings of lasalocid poisoning in buffalo calves. Case: A visit was made to a farm in the municipality of Mojú, Pará state to care for Murrah buffalo calves. After weaning, the buffalos were grazed in paddocks with Panicum spp., and received a supplement of mineral, protein, and vitamin. This supplement contained, per kg, 250 g PB, 50 g Ca, 20 g P, 8 g S, 39 g Na, 20 mg Co, 557 mg Cu, 200 mg Fe, 12.4 mg Se, 2040 mg Zn, 0.19 mg biotin, 26750 IU of vitamin A, 4175 IU of vitamin D, 155 IU of vitamin E and 300 mg/kg of lasalocid. The product was made available to all calves, at 1-2 g/kg body weight (BW), according to the manufacturer's recommendations. Older calves were raised together with those less than 30 days old; as a result, the older calves tended to eat more, which could lead to a supplementation consumption of more than 1 kg body weight per animal per day. It was reported that between 40 and 60 days after the introduction of this supplement, 16 calves fell ill and died due to apathy, motor instability, tremors, and distended neck. The herd had a mortality rate of 33.3%. Two calves underwent a necroscopic examination at the Pathology Section of the Veterinary Institute of the Federal University of Pará. Macroscopic examination revealed extensive pale areas in the skeletal muscles, myocardium, and tongue. Fragments of these muscles and various organs were collected, fixed in 10% buffered formalin, processed according to the routine histological technique, and stained with hematoxylin-eosin and Masson's trichrome stain. Microscopic examination of the histologic samples revealed foci of muscle atrophy and necrosis characterized by an increase in cytoplasmic eosinophilia associated with the loss of stretch marks, and hyperchromatic nuclei that were displaced to the periphery. The necrosis of the muscle fibers was highlighted by Masson's trichrome staining. Discussion: The diagnosis of lasalocid poisoning in buffalo calves was based on epidemiological data, clinical findings, results of macroscopic and histopathological examination, and based on the estimated ionophore intake, obtained directly from the supplement label and by the calf's handler. Based on the absence of stratification of the calves by similarity of age and because the buffalo calves older than 30 days could eat more than 1 kg of the supplement (containing 300 mg/kg of lasalocid), it was possible to estimate the intake of lasalocid per kg CP (body weight). Therefore, the intake of lasalocid by a 70-kg buffalo calf in approximately 90 days and daily supplement consumption between 1 and 1.5 kg would be between 4.2 and 6.4 mg/kg of body weight. This report reinforces that notion that buffalo calves should never ingest ionophores; however, if necessary, strict protocols must be followed to avoid poisoning in these animals. This study highlighted the fact that stratification of buffaloes by different age groups during feeding became a risk factor that allowed greater consumption by older animals; this led to the estimated consumption of 4.2-6.4 mg/kg of lasalocid.

Principais aspectos de listeria monocytogenes e sua importância para a saúde pública / Main aspects of l. Monocytogenes and its importance to public health

Esta revisão tem como objetivo descrever os principais aspectos relacionados a Listeria monocytogenes e sua ocorrência nos alimentos. Alimentos contaminados com L. monocytogenes representam importante risco para o desenvolvimento das doenças veiculadas por alimentos (DVA). Listeriose é uma doença grave para grupos de risco, como idosos, mulheres grávidas, recém-nascidos e pacientes imunocomprometidos, com taxas de letalidade que podem ultrapassar 80%. É um patógeno amplamente distribuído na natureza, pode ser isolado de diversos produtos de origem animal e vegetal, como carnes de frango, bovinos, suínos, embutidos, leite e derivados e hortaliças. Além disso, possui inúmeros fatores de virulência, que permitem que escape do sistema imune e mantenha a infecção. É um microrganismo capaz de crescer em temperaturas de refrigeração e formar biofilme, sendo esses fatores de risco quando associados às más práticas de manejo e controle da cadeia produtiva dos alimentos, já que estas características propiciam a contaminação cruzada. Como medidas de controle, faz-se necessário implementar métodos que assegurem as boas condições de manipulação dos alimentos na indústria, no varejo e nas residências. Muitas práticas previstas na legislação podem ser empregadas para controlar contaminação nos alimentos, como as Boas Práticas de Fabricação (BPF), os Procedimentos Padrão de Higiene Operacional (PPHO) e o sistema de Análise de Perigos e Pontos Críticos de Controle (APPCC). Porém, a falta de fiscalização pelos órgãos competentes pode ser um fator de incentivo para a não implementação destas práticas, o que pode comprometer a qualidade microbiológica dos alimentos e gerar riscos à saúde pública.

This review aims to list the main aspects of Listeria monocytogenes and its occurrence in foods. Food contaminated with L. monocytogenes poses an important risk for the occurrence of foodborne illnesses. Listeriosis is a serious disease to risk groups, such as the elderly, pregnant women, newborns and immunocompromised patients, with mortality rates that can exceed 80%. It is a widely distributed pathogen in the environment and can be isolated from various animal and greens, such as chicken, beef, pork, sausages, dairy products and vegetables. In addition, the pathogen has numerous of virulence factors, that allows the microorganism to dodge the immune system and continue the infection. It is a microorganism capable of growing at refrigeration temperatures and forming biofilm, which is a risk factor when combined with poor management and control practices in the food production chain, as these features can lead to cross contamination. As control measures, it is necessary to implement methods that ensure good food handling conditions in industry, retail and households. Many practices provided in the legislation can be used to control food contamination, such as Good Manufacturing Practices (GMP), Standard Operating Hygiene Procedures (SOHP) and the Hazard Analysis and Critical Control Points (HACCP) system; however, the lack of inspection by competent bodies can be an encouragement factor for the non-implementation of these practices, which would be able to compromise the microbiological quality of the food and creates a threat to public health.

Principais aspectos de listeria monocytogenes e sua importância para a saúde pública / Main aspects of l. Monocytogenes and its importance to public health

Esta revisão tem como objetivo descrever os principais aspectos relacionados a Listeria monocytogenes e sua ocorrência nos alimentos. Alimentos contaminados com L. monocytogenes representam importante risco para o desenvolvimento das doenças veiculadas por alimentos (DVA). Listeriose é uma doença grave para grupos de risco, como idosos, mulheres grávidas, recém-nascidos e pacientes imunocomprometidos, com taxas de letalidade que podem ultrapassar 80%. É um patógeno amplamente distribuído na natureza, pode ser isolado de diversos produtos de origem animal e vegetal, como carnes de frango, bovinos, suínos, embutidos, leite e derivados e hortaliças. Além disso, possui inúmeros fatores de virulência, que permitem que escape do sistema imune e mantenha a infecção. É um microrganismo capaz de crescer em temperaturas de refrigeração e formar biofilme, sendo esses fatores de risco quando associados às más práticas de manejo e controle da cadeia produtiva dos alimentos, já que estas características propiciam a contaminação cruzada. Como medidas de controle, faz-se necessário implementar métodos que assegurem as boas condições de manipulação dos alimentos na indústria, no varejo e nas residências. Muitas práticas previstas na legislação podem ser empregadas para controlar contaminação nos alimentos, como as Boas Práticas de Fabricação (BPF), os Procedimentos Padrão de Higiene Operacional (PPHO) e o sistema de Análise de Perigos e Pontos Críticos de Controle (APPCC). Porém, a falta de fiscalização pelos órgãos competentes pode ser um fator de incentivo para a não implementação destas práticas, o que pode comprometer a qualidade microbiológica dos alimentos e gerar riscos à saúde pública.(AU)

This review aims to list the main aspects of Listeria monocytogenes and its occurrence in foods. Food contaminated with L. monocytogenes poses an important risk for the occurrence of foodborne illnesses. Listeriosis is a serious disease to risk groups, such as the elderly, pregnant women, newborns and immunocompromised patients, with mortality rates that can exceed 80%. It is a widely distributed pathogen in the environment and can be isolated from various animal and greens, such as chicken, beef, pork, sausages, dairy products and vegetables. In addition, the pathogen has numerous of virulence factors, that allows the microorganism to dodge the immune system and continue the infection. It is a microorganism capable of growing at refrigeration temperatures and forming biofilm, which is a risk factor when combined with poor management and control practices in the food production chain, as these features can lead to cross contamination. As control measures, it is necessary to implement methods that ensure good food handling conditions in industry, retail and households. Many practices provided in the legislation can be used to control food contamination, such as Good Manufacturing Practices (GMP), Standard Operating Hygiene Procedures (SOHP) and the Hazard Analysis and Critical Control Points (HACCP) system; however, the lack of inspection by competent bodies can be an encouragement factor for the non-implementation of these practices, which would be able to compromise the microbiological quality of the food and creates a threat to public health.(AU)

Genotypical characterization of thermotolerant coliforms isolated from food produced by a Solidarity Economic Venture of Bahia (Brazil) / Caracterização genotípica de coliformes termotolerantes isolados de alimentos produzidos por um Empreendimento Econômico Solidário

Many Solidarity Economic Venture (SEV) are family farmers who seek to add value to production through artisanal processing, which can lead to food contamination. Thus, this study aimed to genotypically characterize thermotolerant coliforms (TtC) strains from food produced by local agribusinesses of SEV during January to April 2019. Samples from thirteen production units (PU) from the SEV were submitted to a microbiological analysis of thermotolerant coliforms (AFNOR 3M1/2 09/89), using a fast count method in Petrifilm dishes. The Polymerase Chain Reaction (PCR) technique was used to verify the following virulence genes (VGs) associated with Escherichia coli: stx, typical from enterohemorrhagic E. coli (EHEC); bfpA typical from entheropathogenic E. coli (EPEC) and elt and slt, typical from entherotoxigenic E. coli (ETEC). The results showed that two samples of queijadinha (typical Brazilian candy made with eggs and coconut) and one sample of cassava cake presented characteristic colonies TtC. This way, three strains were isolated in order to perform the PCR technique. However, the genes used in the reaction were not detected in the isolated strains. Therefore, it is suggested that the isolated strains are from E. coli pathotypes with different virulence genes than the ones analyzed belong other types of TtC, such as Enterobacter and Klebsiella. Although the virulence of genes has not been confirmed, the presence of TtC on food indicates hygiene flaws during production and, therefore, measurements to control and prevent contamination should be taken.(AU)

Muitos Empreendimentos Econômicos Solidários (EES) são formados por agricultores familiares que buscam agregar valor à produção por meio do beneficiamento artesanal, que pode ocasionar a contaminação dos alimentos. Desta forma, este estudo objetivou caracterizar genotipicamente coliformes termotolerantes (CT) isolados em alimentos produzidos por agroindústrias de um EES no período de janeiro a abril de 2019. Então, foi realizada análise microbiológica de coliformes termotolerantes (AFNOR 3M1/2 09/89), utilizando um método contagem de contagem rápida em placas Petrifilm, em amostras de alimentos de treze Unidades de Produção (UP) do EES. Foram coletadas assepticamente cinco amostras de cada UP, totalizando 65 amostras. Utilizou-se a técnica de Reação em Cadeia de Polimerase (PCR) para verificação dos seguintes genes de virulência de Escherichia coli: stx, característico de E. coli enterohemorrágica (EHEC), bfpA, característico de E. coli enteropatogênica (EPEC) e elt e stI, característicos de E. coli enterotoxigênica (ETEC). Os resultados demonstraram que duas amostras de queijadinha e uma amostra do bolo de aipim apresentaram colônias características de coliformes termotolerantes. Desta forma, foram isoladas três cepas para a realização da PCR, no entanto os genes utilizados nas reações não foram identificados nas cepas isoladas. Portanto, sugere-se que as cepas isoladas sejam de patótipos de E. coli com genes de virulência diferentes dos analisados ou de outro membro dos CT, como Enterobacter e Klebsiella. Apesar de não serem confirmados os genes de virulência analisados, a detecção dos CT nos alimentos indica falhas na higiene durante a produção, portanto medidas para controlar e prevenir a contaminação dos produtos devem ser tomadas.(AU)

Eventração em Iguana verde (Iguana iguana) / Eventration in green iguana (Iguana iguana)

Background: The reptile class could be considered one of the biggest vertebrate groups and are divided in orders andsuborders according to their characteristics. These animals’ maintenance in captivity, either at home, captive bred or atzoos, can generate risk to their health, if the required cares are not given for each respective species. The lack of individualcares could lead to bone and muscular diseases and to traumatic lesions in soft tissues, mainly in the coelomic cavity. Thereport that is being presented aims to describe the case of a green iguana (Iguana iguana) that presented an increase ofvolume in the coelomic cavity. The animal belongs to the squad of the Zoo “Dr. Fábio de Sá Barreto”.Case: A green iguana arrived at the Zoo in February 2019 coming from another establishment, with already an increaseof volume in the coelomic cavity. The animal was put in quarantine and later on, it was put in display at a terrarium in theZoo considered adequate to reptiles, with another seven green iguanas along with an argentine tegu (Salvator rufescens).Their feed was offered in the morning and was composed of fruits, vegetables and flowers like hibiscus. In the end of June2019, it was reported by the attendant that the animal was presented with anorexia and prostration, and these symptomsprogressed to neurologic signs, as it walked in circles. So, the animal was evaluated by the Zoo veterinarians and on examthey noticed lethargy, dehydration, absence of reflexes (pupillary, eyelid and painful), locomotion difficulty and when theiguana moves, it walks in circles. The increase in volume had the same size as reported in February and a soft consistency.After that, the animal was interned and treated according to the symptoms and the clinical evolution. Ten days after thehospitalization, the animal was still not eating, and locomotion stopped completely. It was performed in an...

Eventração em Iguana verde (Iguana iguana) / Eventration in green iguana (Iguana iguana)

Background: The reptile class could be considered one of the biggest vertebrate groups and are divided in orders andsuborders according to their characteristics. These animals maintenance in captivity, either at home, captive bred or atzoos, can generate risk to their health, if the required cares are not given for each respective species. The lack of individualcares could lead to bone and muscular diseases and to traumatic lesions in soft tissues, mainly in the coelomic cavity. Thereport that is being presented aims to describe the case of a green iguana (Iguana iguana) that presented an increase ofvolume in the coelomic cavity. The animal belongs to the squad of the Zoo “Dr. Fábio de Sá Barreto”.Case: A green iguana arrived at the Zoo in February 2019 coming from another establishment, with already an increaseof volume in the coelomic cavity. The animal was put in quarantine and later on, it was put in display at a terrarium in theZoo considered adequate to reptiles, with another seven green iguanas along with an argentine tegu (Salvator rufescens).Their feed was offered in the morning and was composed of fruits, vegetables and flowers like hibiscus. In the end of June2019, it was reported by the attendant that the animal was presented with anorexia and prostration, and these symptomsprogressed to neurologic signs, as it walked in circles. So, the animal was evaluated by the Zoo veterinarians and on examthey noticed lethargy, dehydration, absence of reflexes (pupillary, eyelid and painful), locomotion difficulty and when theiguana moves, it walks in circles. The increase in volume had the same size as reported in February and a soft consistency.After that, the animal was interned and treated according to the symptoms and the clinical evolution. Ten days after thehospitalization, the animal was still not eating, and locomotion stopped completely. It was performed in an...(AU)

Enterococcus Spp. Survival Through the Use of Standard Protocol for Clostridium Sp. Isolation

The genera Clostridium and Enterococcus are very different from each other, both morphologically and physiologically. Due to the high resistance by the sporulation capacity of Clostridium species, the thermal shock is a characteristic tool used for the isolation and identification of these microorganisms, this way, it would eliminate any other bacteria that did not present spores. The objective of this work is to show that Enterococcus sp. resist the temperature treatment and grow in culture media used for the isolation of Clostridium sp. For this, the present study initially attempted to identify reducing sulfite clostridia in poultry products, through the use of specific culture media and heat shock treatment. However, the PCR did not detect the presence of Clostridium sp. Then, sequencing of the 16S rDNA region was performed, which showed that the reducing sulfite colonies that were being isolated were, actually, Enterococcus spp. With this, some tests were carried out using different temperature and time combinations in the thermal shock, as well as the use of five different selective and differential culture media, in an attempt to eliminate any contaminants, but all without success, because these bacteria resisted to all modification. Therefore, the standard protocol for the isolation of bacteria of the genus Clostridium does not eliminate Enterococcus, which can lead to failures in the quantification and qualification of sulfite reducing microorganisms, a fact that can significantly affect food safety and animal health.(AU)

Enterococcus Spp. Survival Through the Use of Standard Protocol for Clostridium Sp. Isolation

The genera Clostridium and Enterococcus are very different from each other, both morphologically and physiologically. Due to the high resistance by the sporulation capacity of Clostridium species, the thermal shock is a characteristic tool used for the isolation and identification of these microorganisms, this way, it would eliminate any other bacteria that did not present spores. The objective of this work is to show that Enterococcus sp. resist the temperature treatment and grow in culture media used for the isolation of Clostridium sp. For this, the present study initially attempted to identify reducing sulfite clostridia in poultry products, through the use of specific culture media and heat shock treatment. However, the PCR did not detect the presence of Clostridium sp. Then, sequencing of the 16S rDNA region was performed, which showed that the reducing sulfite colonies that were being isolated were, actually, Enterococcus spp. With this, some tests were carried out using different temperature and time combinations in the thermal shock, as well as the use of five different selective and differential culture media, in an attempt to eliminate any contaminants, but all without success, because these bacteria resisted to all modification. Therefore, the standard protocol for the isolation of bacteria of the genus Clostridium does not eliminate Enterococcus, which can lead to failures in the quantification and qualification of sulfite reducing microorganisms, a fact that can significantly affect food safety and animal health.

Immunological and biossensory techniques for detection of Salmonella spp. in food derived from fish farming- review / Técnicas imunológicas e biossensoriais para detecção de Salmonella spp. em alimentos derivados da piscicultura ­ revisão de literatura / Técnicas inmunológicas y biosensoriales para detección de Salmonella spp. en alimentos derivados de piscicultura - revisión de literatura

Salmonellosis is the world's most common foodborne illness. In Brazil, foods contaminated by salmonella lead the statistics. Therefore, the aim of this study is, through biotechnological knowledge, to compile alternative and innovative techniques for the detection of salmonella in foods, such as fish-farming derivatives, immunological and biosensorial techniques. This is a descriptive exploratory data survey of a qualitative nature, aiming at data analysis. Research and data collection were carried out in bibliographic databases: Academic Google, Scielo, CAPES journals and institutional repositories using specific descriptors - in Portuguese and English, with words and terms separated by the Boolean operators 'AND' and 'OR'. Some innovative and alternative methods are available to identify the presence of salmonella in food. Immunological and biosensory techniques, despite being less frequent in the scientific literature than molecular methods, are techniques that present high specificity and sensitivity. These techniques have been the most developed alternative methods in fish in recent years. And, they can employ both molecular and immunological techniques in biorecognition, which is characterized as an advantage of not having a requirement for pre-enrichment of the sample. According to the literature found, the techniques covered in this study are quick to respond, which speeds up decision-making by researchers and technicians, which makes the techniques very promising for industrial application.(AU)

A salmonelose é uma enfermidade de maior ocorrência no mundo veiculada por alimentos. No Brasil, alimentos contaminados por salmonelas lideram as estatísticas. Por isso, o objetivo desse estudo é através dos conhecimentos biotecnológicos compilar técnicas alternativas e inovadoras para a detecção de salmonelas em alimentos, como os derivados da piscicultura, as técnicas imunológicas e biossensoriais. Trata-se de um estudo de levantamento de dados descritivo exploratório de de caráter qualitativo, visando à análise dos dados. As pesquisas e coletas de dados foram realizadas nas bases bibliográficas: Google Acadêmico, Scielo, periódicos da CAPES e repositórios institucionais utilizando os descritores específicos - nos idiomas português e inglês, com palavras e termos separados pelos operadores booleanos 'AND' e 'OR'. São disponibilizados alguns métodos inovadores e alternativos para identificação da presença de salmonelas em alimentos. As técnicas imunológicas e biossensoriais, apesar de serem menos frequentes na literatura científica do que os métodos moleculares são técnicas que apresentaram elevada especificidade e sensibilidade. Essas técnicas têm sido os métodos alternativos mais desenvolvidos em peixes nos últimos anos. E, podem empregar tanto técnicas moleculares como imunológicas no biorreconhecimento, o que se caracteriza como vantagem de não haver requerimento de pré-enriquecimento da amostra. Conforme a literatura encontrada, as técnicas abordadas por esse estudo apresentam rapidez de resposta o que agiliza as tomadas de decisões dos pesquisadores e técnicos, o que torna as técnicas bastante promissora para aplicação industrial.(AU)

La salmonelosis es la enfermedad transmitida por alimentos más común del mundo. En Brasil, los alimentos contaminados por salmonelas lideran las estadísticas. Por tanto, el objetivo de ese estudio fue a través de conocimientos biotecnológicos recopilar técnicas alternativas e innovadoras para la detección de salmonelas en los alimentos, como los derivados de la piscicultura, las técnicas inmunológicas y biosensoriales. Se trata de una encuesta de datos exploratorio descriptivo de carácter cualitativo, cuyo objetivo es el análisis de datos. Las investigaciones y recopilaciones de datos se realizaron en bases de datos bibliográficas: Google Académico, Scielo, revistas CAPES y repositorios institucionales utilizando descriptores específicos, en portugués e inglés, con palabras y términos separados por los operadores booleanos 'AND' y 'OR'. Se encuentran disponibles algunos métodos innovadores y alternativos para identificar la presencia de salmonela en los alimentos. Las técnicas inmunológicas y biosensoriales, a pesar de ser menos frecuentes en la literatura científica que los métodos moleculares, son técnicas de alta especificidad y sensibilidad. Esas técnicas han sido los métodos alternativos más desarrollados en peces en los últimos años. Y pueden emplear técnicas tanto moleculares como inmunológicas en el biorreconocimiento, que se caracteriza por la ventaja de no tener un requisito de preenriquecimiento de la muestra. Según la literatura encontrada, las técnicas abordadas en este estudio son de rápida respuesta, lo que agiliza la toma de decisiones por parte de investigadores y técnicos, lo que hace que las técnicas sean muy prometedoras para la aplicación industrial.(AU)

Immunological and biossensory techniques for detection of Salmonella spp. in food derived from fish farming- review / Técnicas imunológicas e biossensoriais para detecção de Salmonella spp. em alimentos derivados da piscicultura ­ revisão de literatura / Técnicas inmunológicas y biosensoriales para detección de Salmonella spp. en alimentos derivados de piscicultura - revisión de literatura

Salmonellosis is the world's most common foodborne illness. In Brazil, foods contaminated by salmonella lead the statistics. Therefore, the aim of this study is, through biotechnological knowledge, to compile alternative and innovative techniques for the detection of salmonella in foods, such as fish-farming derivatives, immunological and biosensorial techniques. This is a descriptive exploratory data survey of a qualitative nature, aiming at data analysis. Research and data collection were carried out in bibliographic databases: Academic Google, Scielo, CAPES journals and institutional repositories using specific descriptors - in Portuguese and English, with words and terms separated by the Boolean operators 'AND' and 'OR'. Some innovative and alternative methods are available to identify the presence of salmonella in food. Immunological and biosensory techniques, despite being less frequent in the scientific literature than molecular methods, are techniques that present high specificity and sensitivity. These techniques have been the most developed alternative methods in fish in recent years. And, they can employ both molecular and immunological techniques in biorecognition, which is characterized as an advantage of not having a requirement for pre-enrichment of the sample. According to the literature found, the techniques covered in this study are quick to respond, which speeds up decision-making by researchers and technicians, which makes the techniques very promising for industrial application.(AU)

A salmonelose é uma enfermidade de maior ocorrência no mundo veiculada por alimentos. No Brasil, alimentos contaminados por salmonelas lideram as estatísticas. Por isso, o objetivo desse estudo é através dos conhecimentos biotecnológicos compilar técnicas alternativas e inovadoras para a detecção de salmonelas em alimentos, como os derivados da piscicultura, as técnicas imunológicas e biossensoriais. Trata-se de um estudo de levantamento de dados descritivo exploratório de de caráter qualitativo, visando à análise dos dados. As pesquisas e coletas de dados foram realizadas nas bases bibliográficas: Google Acadêmico, Scielo, periódicos da CAPES e repositórios institucionais utilizando os descritores específicos - nos idiomas português e inglês, com palavras e termos separados pelos operadores booleanos 'AND' e 'OR'. São disponibilizados alguns métodos inovadores e alternativos para identificação da presença de salmonelas em alimentos. As técnicas imunológicas e biossensoriais, apesar de serem menos frequentes na literatura científica do que os métodos moleculares são técnicas que apresentaram elevada especificidade e sensibilidade. Essas técnicas têm sido os métodos alternativos mais desenvolvidos em peixes nos últimos anos. E, podem empregar tanto técnicas moleculares como imunológicas no biorreconhecimento, o que se caracteriza como vantagem de não haver requerimento de pré-enriquecimento da amostra. Conforme a literatura encontrada, as técnicas abordadas por esse estudo apresentam rapidez de resposta o que agiliza as tomadas de decisões dos pesquisadores e técnicos, o que torna as técnicas bastante promissora para aplicação industrial.(AU)

La salmonelosis es la enfermedad transmitida por alimentos más común del mundo. En Brasil, los alimentos contaminados por salmonelas lideran las estadísticas. Por tanto, el objetivo de ese estudio fue a través de conocimientos biotecnológicos recopilar técnicas alternativas e innovadoras para la detección de salmonelas en los alimentos, como los derivados de la piscicultura, las técnicas inmunológicas y biosensoriales. Se trata de una encuesta de datos exploratorio descriptivo de carácter cualitativo, cuyo objetivo es el análisis de datos. Las investigaciones y recopilaciones de datos se realizaron en bases de datos bibliográficas: Google Académico, Scielo, revistas CAPES y repositorios institucionales utilizando descriptores específicos, en portugués e inglés, con palabras y términos separados por los operadores booleanos 'AND' y 'OR'. Se encuentran disponibles algunos métodos innovadores y alternativos para identificar la presencia de salmonela en los alimentos. Las técnicas inmunológicas y biosensoriales, a pesar de ser menos frecuentes en la literatura científica que los métodos moleculares, son técnicas de alta especificidad y sensibilidad. Esas técnicas han sido los métodos alternativos más desarrollados en peces en los últimos años. Y pueden emplear técnicas tanto moleculares como inmunológicas en el biorreconocimiento, que se caracteriza por la ventaja de no tener un requisito de preenriquecimiento de la muestra. Según la literatura encontrada, las técnicas abordadas en este estudio son de rápida respuesta, lo que agiliza la toma de decisiones por parte de investigadores y técnicos, lo que hace que las técnicas sean muy prometedoras para la aplicación industrial.(AU)

Melanose talâmica em caprinos / Thalamic melanosis in goats

Background: Melanosis is a blackened pigmentation resulting from the accumulation of melanocytes in tissues that are not normally pigmented. This change in the color of the organs occurs due to the agglomeration of melanocytes originating from abnormal migration during embryogenesis and does not cause dysfunction to the affected organ. Although melanosis frequently occurs in several species and affects several organs such as the brain and spinal cord leptomeninges, involvement in the thalamus region is unusual. The objective of this work was to report 2 cases of thalamic melanosis in goats, determining the pathological and histochemical aspects that assist in the diagnosis of this condition. Cases: Two cases of thalamic melanosis in goats were diagnosed. In both cases, the animals had no nervous history disease and clinical signs. The cause of death in cases 1 and 2 was established based on anatomopathological findings and clinical signs being diagnosed with mycoplasmosis and asphyxia, respectively. After fixing and making cross-sections of the brain, a focal lengthy blackened area was observed on the thalamus surface in both cases. Microscopically, lesions in the brain were similar in both cases and exclusively affected the thalamus. These cells had abundant cytoplasm, well delimited with brownish granular pigment. The nuclei were difficult to visualize and in some cells, it was rounded, well-defined, morphologically compatible with melanocytes. Melanocytes were mainly distributed around neurons and often distended the perivascular space of multiple blood vessels. In Fontana Masson staining, the granules in the cytoplasm of these cells stained strongly black. The Prussian Blue, Periodic Acid- Schiff's, Von Kossa, and Giemsa stains were negative, and the pigment remained brown. In the unstained slides, assembled after the deparaffinization and clarification process, it was observed the permanence of cells with blackish-brown pigment in the cytoplasm. In immunohistochemistry, strong immunostaining of pigmented cells with the Anti-MelanA antibodies was observed in both cases. Discussion: The diagnosis of thalamic melanosis in goats was carried out based on the characteristic pathological findings, in which melanin pigments were demonstrated and identified through HE, Fontana-Masson staining, and unstained slides and confirmed by the IHC. The use of complementary histochemical techniques was fundamental for the classification of the pigment as melanin, demonstrating to be an accessible and reliable tool for the diagnosis of pathological processes that lead to the accumulation of pigments and or material in the tissues. The occurrence of melanin in the thalamus may be associated with a failure in the migration of melanoblasts, which would go to the optical pathways or to the thalamus. This erratic migration of melanoblasts can be explained by the fact that the forebrain is the embryogenic origin of the optic and diencephalon pathways. Macroscopically, thalamic melanosis must be differentiated mainly from neoplastic processes such as melanoma and hemangiosarcoma, pigmented fungus infections, Phalaris angusta poisoning, listeriosis, neurocutaneous melanosis, and neuromelanin. It was concluded that thalamic melanosis is an uncommon alteration in goats and although it has been diagnosed as an incidental necropsy finding, should be included in the differential diagnosis of diseases that affect the central nervous system, especially those that have a color change associated with the deposition of pigments in the tissues.

Biosorption potential of lead tolerant fungi isolated from refuse dumpsite soil in Nigeria

Metals are non-biodegradable and recurrent in the environs. Heavy metals tolerant fungiwere isolated from refuse dumpsite soil using pour plate method. These fungiwere identified as Aspergillus niger, Penicillium chrysogenumandRhizomucor sp. The fungal isolates were screened for cadmium (Cd), lead (Pb) and zinc (Zn) with concentration of 200ppm, 400ppm and 600ppm. Aspergillus nigerand Penicillium chrysogenumshowed high tolerance for the metals in contrast to the control. The fungiwith high tolerance were used for biosorption study. However, Penicillium chrysogenumshowed higher lead removal or biosorption potential of 1.07ppm, 3.35ppm and 4.19ppm as compared with Aspergillus nigerwith lead removal of 0.67ppm, 3.11ppm and 3.79ppm at 5th, 10thand 15thday respectively. One-way Analysis of Variance was used to interpret the data generated from the biosorption study which revealed that there was no significant different (p>0.05)between the lead removal of Aspergillus nigerandPenicillium chrysogenumon the 5thday but there was significant difference (p<0.05)in the lead removal of Aspergillus nigerand Penicillium chrysogenumon the 10thand 15thday. This study suggests the use of these fungal isolates for removal and biotreatment of heavy metal contaminated and polluted environment.

Analysis of NH3-N Slow Release systems for fiber digestibility of low-quality forage: in vitro approach

Urea is a common non-protein supplement used in ruminant feed; however, excessive consumption may lead to poisoning by NH3. Although the slow release of urea into the rumen has shown to be an essential aspect for ruminant feed, to date only a few studies have addressed this matter. In this study we examined the influence of five different NH3-N slow release systems based on clay-urea nanocomposites on the fiber digestibility of low-quality forage (sugarcane straw) in vitro. Physical properties of nanocomposites were evaluated and their effects on digestibility were tested in vitro using pristine urea as a positive control (level of 1 % of DM of sugarcane straw sample) and sugarcane (with no additives) as a negative control. Ammonia release and digestibility were evaluated at 12, 24, 36, 48, 72 and 96-h. Generally, all nanocomposites increased (p < 0.05) digestibility of fiber over control under all the conditions stipulated, but the samples with hydrogel content were more expressive. We concluded that an ideal release rate and optimum environment for microbial synthesis are necessary to maximize the digestion of sugarcane.