TLC and HPLC methods for the determination of plumbagin for the diagnosis of poisoning by Plumbago scandens L.

Plumbago scandens L. (Plumbaginaceae) occurs in all regions of Brazil. It has been described as toxic to cattle and goats. Caustic lesions in the upper digestive tract characterize poisoning. P. scandens contains a naphthoquinone named plumbagin, which presents high cytotoxic activity. Plumbago auriculata Lam., a widely used ornamental plant, is considered potentially toxic, but there is limited data about its toxicity. This work aimed to validate analytical methodologies for determining the levels of plumbagin in samples of leaves, stems, and rumen content to be used as an auxiliary chemical marker in the laboratory diagnosis of intoxication. One methodology used thin layer chromatography (TLC), and another used high-performance liquid chromatography (HPLC). The presence of palisade grass (Urochloa brizantha (Hochst. ex A.Rich.) R.D.Webster), Guinea grass (Megathyrsus maximus (Jacq.) B.K.Simon & S.W.L.Jacobs), corn silage, and rumen content did not interfere with plumbagin in the two methodologies. The TLC methodology generates qualitative results but is simple to implement and has a low cost. The HPLC methodology showed a limit of detection (LOD) of 0.01 µg/mL and a limit of quantification (LOQ) of 0.05 µg/mL. Leaf and stem samples of P. scandens evaluated showed high levels of plumbagin (0.261 ± 0.087 % and 0.327 ± 0.055 %, respectively). In contrast, leaves of P. auriculata did not show detectable levels of the toxin, and some stem samples showed low levels (up to 0.000114 %). Thus, these methodologies can be used to confirm or rule out the consumption of P. scandens in rumen content from animals suspected of poisoning.

The Health Status of Horses Used for at Least Six Complete Cycles of Loxoscelic Antivenom Production.

Antivenom production against Loxosceles venom relies on horses being immunized and bled for plasma harvest. One horse can partake in several cycles of antivenom production, which will require years of constant venom and adjuvant inoculation and bleeding. The actual impact on the health of horses that participate in several antivenom-producing cycles is unknown. Therefore, this study aimed to evaluate the general health status of horses that underwent at least six cycles of loxoscelic antivenom production. Seven crossbred horses that had partaken in six to eight complete antivenom-producing cycles were used and established as the immunized group (IG). Under the same handling and general management, eleven horses were established as the control group (CG). The horses were evaluated regarding their general clinical status and had their blood sampled, and an ECG recorded. The IG presented lower RBC and PCV, despite keeping values within inferior limits for the species. Renal function was not impaired, and liver-related enzymes were higher than those in the CG, probably due to liver exertion from immunoglobulin synthesis. ECG showed some abnormalities in the IG, such as atrioventricular block and a wandering atrial pacemaker, corroborated by an increase in CK-MB. The cardiovascular abnormalities were mainly found in the horses that participated in several antivenom-producing cycles. The overall results indicate that these horses had some impairment of their general health status. Once available, some alternative, less toxic antigens should replace the venom for immunization of horses used for antivenom production.

Toxic plants from the perspective of a "Quilombola" community in the Cerrado region of Brazil.

A multi-disciplinary team surveyed ranchers at the Kalunga Historical and Cultural Heritage Site, in the Cerrado region of west central Brazil, to determine impacts promoted by toxic plants on cattle. The expedition to the Kalunga region was carried out by Brazilian and American researchers. Previously selected cattle ranch properties from "Vão das Almas", "Engenho II" and "Vão do Moleque" were visited. Twenty-four interviews were carried out with cattle ranchers and a questionnaire was applied to obtain information about outbreaks of native plant poisoning and their effects on livestock, and the use of local plants in phytotherapy. We classified problematic plants into three distinct categories. First, the toxic plants most cited by residents causing cattle losses were the flowers of Caryocar brasiliense Cambess ("pequi"), the fruits of Terminalia corrugata (Ducke) Gere & Boatwr. (Buchenavia tomentosa Eichler - "mirindiba" or "pau-pilão"), Eugenia dysenterica (Mart.) DC ("cagaita"), and Palicourea marcgravii A. St. Hil ("erva-café" or "cafezinho"). Secondly, other plants considered toxic, but causing less severe losses were Emmotum nitens (Benth.) Miers ("casco d'anta"), Indigofera lespedezioides (Kunth) ("timbozinho"), Ricinus communis L. ("mamona"), Pteridium esculentum (G. Forst.) Cockayne ("samambaia"), Stryphnodendron adstringens (Mart.) Coville ("barbatimão"), and Actinocladum verticillatum (Nees) McClure ex Soderstr. ("cambaúba"). The most important finding was the identification of the C. brasiliense flower as potentially toxic to cattle, which must be subject for future research. Further, we confirmed the toxicity and importance of P. marcgravii, E. dysenterica, and Terminalia corrugata. The survey highlighted phytotherapy plants used by the community, and greatly increased awareness by local livestock producers of poisonous plants for management purposes. We conclude that ethnobotanical knowledge, especially from the traditional community, is essential to understand livestock losses to toxic plants, and should be valued not only for reducing livestock losses, but also for cultural importance to the Kalunga communities in the Cerrado.

Influence of landscape ecology and physiological implications in bats from different trophic guilds.

Bats may serve as bioindicators of human impact on landscape ecology. This study aimed to evaluate the health condition of bats from different food guilds captured in two areas with different land use profiles in Brazil and to compare data on the oxidant-antioxidant balance and histopathological changes due to different anthropogenic pressures. Bats were collected from a protected area in Serra do Cipó National Park (SCNP), MG, Brazil, and an area with intense agricultural activity in the municipality of Uberaba (UB), MG, Brazil. Despite the differences in land use and occupation between the studied areas, bats showed similar responses. However, the trophic guilds were affected differently. Frugivorous bats in both areas showed lower activities of the enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST) and concentrations of malondialdehyde (MDA) than other guilds, which can be explained by the greater intake of antioxidants from the diet in addition to the lower production of reactive oxygen species (ROS). Histopathological analysis of the livers revealed that the animals had a similar prevalence in the two areas, with some differences related to guilds. Compared with other bats, hematophagous bats from SCNP had a higher prevalence of steatosis and, together with frugivorous bats from Uberaba, had higher frequencies of ballooning degeneration, suggesting that these animals are subjected to anthropogenic factors capable of inducing disturbances in hepatic metabolism. Hematophagous bats from Uberaba had a higher prevalence of portal inflammation, while insectivorous bats from Uberaba had a higher prevalence of lobular and portal inflammation. The profiles of use and occupation of the areas are different; Uberaba bats seem to face worse conditions because they show more liver damage owing to lipoperoxidation.

Cervical Vertebral Stenotic Myelopathy in a Nelore Calf.

This paper aims to report clinical, laboratory, radiographic, and pathological features in a case of cervical vertebral stenotic myelopathy (CVSM) affecting a 4-month-old Nelore calf for the first time. During physical examination, the calf could stand if assisted when lifting by the tail but fallen to the ground when trying to walk. Attempts to flex and extend the neck to the right side failed. Radiographs findings consisted of reduced intervertebral spaces, and misalignments between the endplates, more evident between the C3 and C4 vertebrae, resulting in narrowing of the spinal canal and compression of the spinal cord. Grossly, C4 showed cranial articular surface malformation, abnormal metaphyseal growth plate development, reduced vertebral body size and deformity. Histologically, C4 showed an abnormal vertebral bone development characterized by moderate replacement of trabecular bone by fibrous tissues, multifocal areas of dystrophic hyaline cartilage development, and cartilaginous growth failure along the metaphyseal growth plate. Cervical spinal cord within the stenotic vertebral canal showed swollen neurons with central chromatolysis, areas of Wallerian degeneration, and necrotic debris. In contrast with the well-known Wobbler syndrome in horses, the etiology of CVSM in cattle remains undetermined, and further genetic and pathological studies must be conducted to elucidate it.

Protective Effectiveness of an Immunization Protocol Against the Toxic Effects of Loxosceles intermedia Venom in Rabbits.

Loxosceles spp. (brown spiders) bites are responsible for the development of a syndrome consisting mainly of dermonecrotic lesions, and also systemic effects. Rabbits are one of the main experimental models used for better understanding the systemic and local effects of Loxosceles venom. The aim of this study is to evaluate the toxic and protective effects of rabbits immunized with Loxosceles spp. venom. Male New Zealand rabbits were allocated as a control group (CG; n = 5) that received adjuvant (Montanide) and phosphate-buffer saline (PBS), or as venom group (VG; n = 5) that received 21 µg of Loxosceles venom using Montanide as adjuvant. After five immunization cycles, a trial with 7 µg of Loxosceles intermedia (L. intermedia) venom was performed, and dermonecrotic lesions were measured. The rabbits were then euthanized, and their organs were collected for histopathology analysis. Rabbits that had undergone Loxosceles venom immunization protocol showed minor clinical disturbances during the experimental period. The used immunization protocol protected the rabbits against the toxic effect of the Loxosceles venom because they showed minor clinical disturbances during the experimental period.

Miniaturized QuEChERS extraction method for the detection of multi-residue pesticides in bat muscle tissue.

Habitat loss and fragmentation are among the greatest threats to biodiversity and ecosystem stability, with physiological implications on wild fauna. Bats (Microchiroptera) are small mammals with a wide variety of eating habits, and the well-being of these animals is disturbed by exposure to pesticides. This study aimed to develop a miniaturized QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) extraction method for the detection of multi-residue pesticides in bat muscle tissue using gas chromatography coupled with mass spectrometry (GC-MS). A total of 48 pesticides were tested in 250 mg of bat muscle tissue. The developed analytical method was applied to 148 bats collected from two different areas in Minas Gerais State, Southeast Region of Brazil. The method presented good sensitivity and allowed the determination of residues of 48 pesticides in bat muscle using GC-MS. The miniaturized extraction method makes the analysis feasible even when the sample volume is limited. However, no pesticide residues were detected in bats from the two areas investigated.

Clinical Effects of the Immunization Protocol Using Loxosceles Venom in Naïve Horses.

Bites of brown spiders (Loxosceles spp.) are responsible for dermonecrotic lesions and potentially systemic envenoming that can lead to death. The only effective therapy is the use of the antivenom, usually produced in horses. However, little is known about the consequences of the systematic use of the Loxosceles venom and adjuvants and of the bleedings on antivenom-producing horses. Thus, the aim of this study was to evaluate the clinical changes in horses in their first immunization protocol for Loxosceles antivenom production. Eleven healthy horses, never immunized, were evaluated in three different periods: T0 (before immunization); T1 (after their first venom immunization); and T2 (after their first bleeding). Horses were clinically evaluated, sampled for blood, and underwent electrocardiographic (ECG) recordings. Several suppurated subcutaneous abscesses occurred due to the use of Freund's adjuvants and thrombophlebitis due to systematic venipunctures for the bleeding procedures. ECG showed arrhythmias in few horses in T2, such as an increase in T and R waves. In summary, the immunization protocol impacted on horses' health, especially after bleeding for antivenom procurement.

Neurological and gastrointestinal manifestations of spontaneous poisoning by Ricinus communis in goats.

An outbreak of Ricinus communis poisoning in goats with neurological and digestive changes was related to the ingestion of different vegetative parts of the plant. Two poisoned animals died within 5 h of the plant intake showing necrotic gastroenteritis and hepatocytes degeneration and necrosis. Toxicological analysis by HPLC-DAD assay demonstrated 21.1-25.1 µg/g of ricinine in samples of ruminal fluids and 10.1-10.9 µg/g in the liver of poisoned goats.

Downer cow syndrome causing rhabdomyolysis and acute renal failure in a 17-month-old Guzerá heifer / Síndrome da vaca caída causando rabdomiólise e insuficiência renal aguda em uma novilha Guzerá de 17 meses

The downer cow syndrome (DCS) is characterized by an alert cow showing inability or reluctance to stand for 12 hours or more. This paper reported clinical, laboratory, and pathological findings in a Guzerá heifer with rhabdomyolysis, pigmenturia and acute renal failure following DCS. A 17-month-old Guzerá heifer was transported via a 350-km ride in a truck and showed sternal recumbency and severe difficulty in standing and walking. Neurological examination was unremarkable, and the heifer presented normal response to cranial nerves and spinal cord tests. Rectal palpation revealed a 5-month gravid uterus. No other abnormalities were noted in the pelvis or around the coxofemoral joints. Biochemical abnormalities included extremely high muscular enzyme activities (creatine phosphokinase and aspartate aminotransferase) and high creatinine levels. Urinalysis revealed blackish and cloudy urine, proteinuria, and a positive occult blood test. Spinal cord ultrasonography showed no abnormalities. This report highlighted an uncommon clinical presentation (myoglobinuria) and pathological findings in a heifer with DCS as a consequence of severe compressive muscle damage. Practitioners and producers must be aware of the risk of careless road transportation for long distances of cattle, especially obese cows, avoiding unnecessary suffering and expenses due to DCS.

A síndrome da vaca caída (SVC) é caracterizada por um bovino alerta que mostra incapacidade ou relutância em permanecer em estação por 12 horas ou mais. O objetivo deste trabalho é relatar os achados clínicos, laboratoriais e patológicos em uma novilha Guzerá com rabdomiólise, pigmentúria e insuficiência renal aguda após a SVC. Uma novilha da raça Guzerá, de 17 meses de idade, foi transportada de caminhão por 350 km e apresentou decúbito esternal, grande dificuldade para assumir estação e caminhar. O exame neurológico não demonstrou alterações, e a novilha possuía resposta normal aos testes de nervos cranianos e medula espinhal. A palpação retal revelou útero grávido de cinco meses. Nenhuma outra anormalidade foi observada na pelve ou na região das articulações coxofemorais. As anormalidades bioquímicas incluíram atividades de enzimas musculares (creatina fosfoquinase e aspartato aminotransferase) extremamente aumentadas e níveis elevados de creatinina. A urinálise revelou urina enegrecida e turva, proteinúria e teste de sangue oculto positivo. O exame ultrassonográfico da medula espinhal não apresentou anormalidades. Este relato evidencia uma apresentação clínica (mioglobinúria) e achados patológicos incomuns em uma novilha com SVC em consequência de extensa lesão muscular compressiva. Veterinários e produtores devem estar atentos aos riscos do transporte rodoviário descuidado por longas distâncias de bovinos, especialmente vacas obesas, evitando assim sofrimento e despesas desnecessárias decorrentes da SVC.

Crotalaria spectabilis poisoning in horses fed contaminating oats.

The present report describes the clinical and pathological changes induced by the consumption of oats contaminated with Crotalaria spectabilis seeds by horses. Eighty horses were exposed to oats containing 10 g/kg of C. spectabilis seeds with 0.46% pyrrolizidine alkaloids, and 21 horses died within a 6-month period. Clinical signs included jaundice, apathy, a hypotonic tongue, ataxia, hyporexia, weight loss, aimless wandering, violent behavior, and proprioceptive deficits. Pathological findings were predominant in the liver and included periportal bridging fibrosis, megalocytosis, centrilobular necrosis, and bile stasis. Other findings were Alzheimer's type II astrocytes in the cortex, midbrain, basal nuclei, brainstem and pons; multifocal edema and hemorrhage in the lungs; and degeneration and necrosis of the tubular epithelium of kidneys. Horses are highly sensitive to pyrrolizidine alkaloid-containing plants, and the observed clinical and pathological findings are typical of this poisoning. The seeds were planted, and botanical identification of the adult plants confirmed the diagnosis of C. spectabilis poisoning.

Case Report: A Possible Case of Congenital Erythropoietic Porphyria in a Gir Calf: A Clinical, Pathological, and Molecular Approach.

Congenital erythropoietic porphyria (CEP) is a rare hereditary autosomal recessive disease which has never been reported in Zebu cattle. A 3-day-old Gir calf showed teeth discoloration, fever, dehydration, and dyspnea. The main gross findings were pink-colored teeth, red-brown periosteum and bone marrow, and a fluorescent bright pink coloration of the bone marrow and articular surfaces under ultraviolet light. Aggregates of periodic acid-Schiff (PAS)-stained porphyrin pigments were evident in the lungs, kidneys, and the liver. An intron 8 single-nucleotide polymorphism (SNP) in both the Gir calf and control animals, along with the absence of the uroporphyrin III synthetase (UROS) gene mutation, was observed. Most SNPs were located in the intron regions of the UROS gene without relevance for CEP. A continuous loss of genetic variability and an increase in inbreeding in some herds may be related to CEP in Gir cattle, one of the most prominent Zebu breeds worldwide. In summary, this study describes a presumptive case of CEP in a Gir calf based on clinical and pathological findings. A definitive diagnosis would require the measurement of porphyrin levels in blood, urine, or tissues or the identification of UROS gene defects.

Intravenous lipid emulsion treatment in rabbits with ivermectin toxicosis.

OBJECTIVE: To determine the effect and safety of IV lipid emulsion in rabbits with acute ivermectin toxicosis. DESIGN: Randomized controlled trial. SETTING: University research facility. ANIMALS: Twenty-four healthy male adult New Zealand rabbits. INTERVENTIONS: Three groups of rabbits (IV, IV_RL, and IV_LE) received 80 mg/kg of ivermectin (8 mL/kg) through a nasogastric tube, and 1 group (LE) received an equivalent volume (8 mL/kg) of 0.9% sodium chloride. Group IV_RL was treated with Ringer's lactate (2 mL/kg bolus, followed by 0.25 mL/kg/min for 60 minutes), whereas groups IV_LE and LE received 20% lipid emulsion. The rabbits were submitted to clinical and neurological evaluation, and blood samples were collected for biochemical analysis. All animals were euthanized, and tissue samples were collected and processed for histopathological evaluation and ivermectin quantification. MEASUREMENTS AND MAIN RESULTS: All animals exposed to ivermectin manifested clinical changes consistent with toxicosis, but the ones that received IV lipid emulsion infusion showed no significant clinical improvement. Intense increase in serum glucose and triglyceride concentrations was seen after ivermectin exposure, along with increased urea and creatinine concentrations, but the last 2 remained within the reference range. Lipid emulsion caused an intense increase in triglycerides and cholesterol concentrations. No pathological abnormalities were seen in the organs sampled. Toxicological analysis showed greater ivermectin concentration in adipose tissue and liver, followed by kidney and, finally, brain. The treatments did not change ivermectin tissue concentration. CONCLUSIONS: When given to rabbits intoxicated with ivermectin, IV lipid emulsion was biochemically and histologically safe but was not effective in treating, delaying, or reversing clinical signs and progression, nor did it alter ivermectin tissue concentration.

History, challenges and perspectives on Loxosceles (brown spiders) antivenom production in Brazil.

Antivenom is the only effective therapy for treating any envenomation. Despite its obvious public health importance, the laborious process of procuring, distributing and controlling the quality of such immunobiologicals is being neglected. Brazil is fully self-sufficient in the production of antivenoms. Since the 1950s, Loxoscelism, a syndrome with an onset after a spider bite from specimens of the Loxosceles genus occurs, is considered a public health issue. The Brazilian history in developing antivenom therapy, its production hindrances, and other challenges are discussed in this paper, as well as some promising novelties that can improve production and processing of Loxosceles antivenom.

Hematological and Serum Biochemical Changes and Their Prognostic Value in Horses Spontaneously Poisoned by Crotalaria spectabilis.

Determining the prognosis of poisoning by plants containing pyrrolizidine alkaloids is usually challenging. This study aimed to identify important prognostic parameters that can determine the severity of spontaneous poisoning by Crotalaria spectabilis in horses. Blood samples from 42 horses spontaneously poisoned by oats contaminated with C. spectabilis seeds were evaluated. Complete blood counts (CBC) and serum biochemical tests [urea, creatinine, total protein, albumin, total and direct bilirubin concentrations, aspartate aminotransferase (AST), γ-glutamyl transferase (GGT), and creatine kinase (CK) activities] were performed. Horses were followed up for 12 months to determine the long-term survival rate; after 12 months, they were divided into two groups: survivors (n = 30) and non-survivors (n = 12). Horses spontaneously poisoned with C. spectabilis had higher levels of urea, globulin, bilirubin (total, direct, and indirect), AST, GGT, and CK than the reference values. Non-survivor horses showed significantly higher (p < 0.05) values of hemoglobin, GGT, and direct bilirubin than the survivor horses. Horses with serum GGT activity higher than 95 U/l had 14.0 times the risk of death compared to animals showing activities equal to or lower than this value, whereas horses with serum direct bilirubin concentration higher than 0.6 mg/dl (10.26 µmol/L) had 5.78 times the risk of death compared to the others. In summary, serum GGT activity and direct bilirubin concentration may be useful prognostic indicators for assessing the severity of C. spectabilis-poisoned horses.

Exposure to pesticides in bats.

Bats provide a variety of ecological services that are essential to the integrity of ecosystems. Indiscriminate use of pesticides has been a threat to biodiversity, and the exposure of bats to these xenobiotics is a threat to their populations. This study presents a review of articles regarding the exposure of bats to pesticides published in the period from January 1951 to July 2020, addressing the temporal and geographical distribution of research, the studied species, and the most studied classes of pesticides. The research was concentrated in the 1970s and 1980s, mostly in the Northern Hemisphere, mainly in the USA. Of the total species in the world, only 5% of them have been studied, evaluating predominantly insectivorous species of the Family Vespertilionidae. Insecticides, mainly organochlorines, were the most studied pesticides. Most research was observational, with little information available on the effects of pesticides on natural bat populations. Despite the advances in analytical techniques for detecting contaminants, the number of studies is still insufficient compared to the number of active ingredients used. The effects of pesticides on other guilds and tropical species remain poorly studied. Future research should investigate the effects of pesticides, especially in sublethal doses causing chronic exposure. It is crucial to assess the impact of these substances on other food guilds and investigate how natural populations respond to the exposure to mixtures of pesticides found in the environment.

Traumatic reticuloperitonitis combined with embolic pneumonia and hepatitis as unusual symptoms of foreign body syndrome in a Holstein bull.

Traumatic reticuloperitonitis combined with embolic pneumonia and hepatitis is unusual signs of foreign body syndrome in cattle. A 4-year-old Holstein bull presented decreased appetite, dry cough, progressive weight loss, sternal recumbence and reluctance to stand and move. Laboratory tests revealed leucocytosis (18.4 × 103 /µl) accompanied by neutrophilia (10.48 × 103 /µl), and monocytosis (1.28 × 103 /µl), hyperglobulinaemia (6.3 g/dl), hypoalbuminaemia (1.5 g/dl), hyperfibrinogenaemia (10 g/L) and severe increase in gamma-glutamyl transferase activity (1,216 U/L). Reticular ultrasonographical examination showed a large amount of hyperechoic and hypoechoic content between the reticular serosa and the hepatic visceral surface. The main gross findings included fibrin deposition and adhesions between the reticulum, liver and diaphragm surfaces; a 4.0 mm in diameter transmural reticular perforation; a 12.0-cm diameter and scarce small randomly abscesses in the liver's parenchyma. The lungs presented multifocal areas of suppurative embolic foci (pulmonary abscesses), interstitial emphysema and multifocal fibrin deposition on the pleural surface. Ancillary diagnostic tests, such as ultrasonography and laboratory test, associated with clinical evaluation, may increase the accuracy of the correct diagnosis and avoid wasting time and money on untreatable cases.

Toxicological Risk Assessment of the Accidental Ingestion of a Honeybee (Apis mellifera L.) Present in Food.

The aim of the present work was to evaluate the possible risk of toxic effects due to the ingestion of a honeybee (Apis mellifera L.) accidentally present in food. The methodology used in this study was a bibliographic survey of studies on the toxic effects related to honeybees, with a critical analysis of the possible risks of accidental ingestion of these insects. The amount of venom present in a bee is considered insufficient to induce detectable toxic effects in a person who ingests it by accident, and various components of the venom are destroyed by gastric secretions. However, despite the rare frequency, there is a risk of the ingestion of a bee, causing an allergic reaction to some components of the venom in sensitized individuals. In addition, pollen carried by a bee may cause an allergic reaction in a sensitive individual. Thus, the accidental ingestion of a bee present in a food does not pose the risk of toxic effects for the majority of the population but may promote allergic reactions in susceptible individuals.

Pathological features of cloned calves that died in the neonatal period / Aspectos patológicos de bezerros clonados com óbito no período neonatal

Somatic-cell nuclear transfer is a cloning technique that enables the creation of a viable embryo from a donor adult to produce a genetically identical individual. This technique opens numerous potential possibilities for medicine and animal reproduction. However, several reports have documented cloning-related issues. Embryo and fetal losses remain significantly higher than in other techniques, and there is a high incidence of dystocia and hydrops, which decreases efficiency and increases costs. Animals delivered at term often exhibit a syndrome known as macrosomia and experience difficulties in adapting to life outside the uterus, and death is a common outcome. In the present study, 41 cloned calves that died in the neonatal period were subjected to gross and histopathological examination. Most important gross lesions were found in the liver (enlargement, congestion, yellowish color), kidneys (brownish color at surface and cut, and cysts), lungs (atelectasis, parenchymal consolidation, and secretions in bronchi and bronchioles), and heart (concentric and eccentric hypertrophy, hematic cysts, persistence of ductus arteriosus). Primary microscopic findings were seen in the liver, kidneys, and lungs from neonatal calves. In the liver, 85% of the animals exhibited hepatic degeneration. The presence of a brownish pigment within the cortical tubules of the kidneys was found in approximately 90% of the samples; the presence of this pigment has not been previously reported in cloned calves. In the lungs, a large number of animals exhibiting lesions characteristic of pneumonia (55%). These changes were the pivotal causes of death, mainly due to problems in adapting to life outside the uterus and opportunistic infections in the neonatal period. Further investigation focusing on pathological anatomical changes is necessary to map these abnormalities in cloned animals.(AU)

A transferência nuclear de células somáticas ou clonagem é uma técnica que permite produzir um indivíduo geneticamente igual a um outro indivíduo adulto. Esta técnica abre inúmeras possibilidades para a medicina e para a reprodução animal. Porém, existem inúmeros relatos de problemas associados à clonagem. A taxa de perda nos períodos embrionário e fetal ainda é muito alta quando comparada a outras biotécnicas; além disso, há uma maior incidência de hidropsias e distocias, diminuindo a eficiência e aumentando o custo da técnica. Os animais que vem a termo frequentemente apresentam uma síndrome chamada de macrossomia, e apresentam dificuldades de adaptação à vida extrauterina e, por isso, o óbito é um desfecho comum. No presente trabalho realizou-se necropsia e coleta de fragmentos de órgãos para avaliação histopatológica de 41 bezerros com óbito neonatal. As lesões macroscópicas mais importantes foram encontradas no fígado (hepatomegalia, congestão e coloração amarelada), rins (coloração amarronzada na superfície e ao corte, e cistos), pulmões (atelectasia, parênquima consolidado, e secreções nos brônquios e bronquíolos), e coração (hipertrofia concêntrica e excêntrica, cistos hemáticos e persistência de ducto arterioso). As principais alterações microscópicas observadas foram presença de pigmento acastanhado no interior dos túbulos corticais renais (aproximadamente 90% dos animais), degeneração hepática (85% das amostras avaliadas) e lesões características de pneumonia (55% dos animais). A pigmentação acastanhada no interior dos túbulos corticais é uma alteração que ainda não havia sido relatada anteriormente em animais clonados. As alterações observadas nestes órgãos foram determinantes para o óbito, e devem ter ocorrido sobretudo devido a problemas na adaptação ao ambiente extrauterino e em decorrência de infecções adquiridas no período neonatal. Os achados encontrados no presente trabalho denotam a necessidade de investigação anatomopatológica detalhada de animais clonados inviáveis, na tentativa de mapear as anormalidades apresentadas por eles.(AU)

Effects of Caffeine on Egg Quality and Performance of Laying Hens.

This study's objective was to determine the effects of caffeine intake at various levels, incorporated in the layers' food, on performance and egg quality of hens. A total of 576 hens, aged 56 weeks, were used. The layers were fed rations containing 0 (control), 150, 300, or 450 ppm of caffeine for 12 weeks. During the experimental period, performance parameters (weight, feed consumption, and livability) and egg production and quality (weight, Haugh unit, percentages of yolk, albumen and eggshell, yolk color, eggshell thickness, and resistance, and calcium and phosphorus eggshell contents) were evaluated. The highest concentration of caffeine in the diet (450 ppm) promoted a significant increase in the mortality of hens (1.45% per week) compared to controls (0.23%). There was a reduction in feed consumption by hens, decreased egg production, and reduced eggshell thickness and percentage, with the increase of caffeine. The egg yolk percentage was increased, and the eggshell percentage was reduced in the groups treated with 300 and 450 ppm of caffeine. Furthermore, reduced eggshell thickness was found in all groups that received caffeine. However, it was found that 150 ppm of caffeine in the food did not cause significant changes in most egg production and quality parameters. In summary, caffeine consumption by laying hens increased mortality rate and promoted deleterious effects on chicken production and egg quality at concentrations of 300 and 450 ppm.