Lung Fragments Research Articles

COVID-19 and Pneumocystis jirovecii Pulmonary Coinfection—The First Case Confirmed through Autopsy

Background: Establishing the diagnosis of COVID-19 and Pneumocystisjirovecii pulmonary coinfection is difficult due to clinical and radiological similarities that exist between the two disorders. For the moment, fungal coinfections are underestimated in COVID-19 patients. Case presentation: We report the case of a 52-year-old male patient, who presented to the emergency department for severe dyspnea and died 17 h later. The RT-PCR test performed at his admission was negative for SARS-CoV-2. Retesting of lung fragments collected during autopsy revealed a positive result for SARS-CoV-2. Histopathological examination showed preexisting lesions, due to comorbidities, as well as recent lesions: massive lung thromboses, alveolar exudate rich in foam cells, suprapleural and intra-alveolar Pneumocystis jirovecii cystic forms, and bilateral adrenal hemorrhage. Conclusion: COVID-19 and P. jirovecii coinfection should be considered, particularly in critically ill patients, and we recommend the systematic search for P. jirovecii in respiratory samples.

Porcine lymphotropic herpesvirus (Gammaherpesvirinae) DNA in free-living wild boars (Sus scrofa Linnaeus, 1758) in Brazil.

BackgroundSuid gammaherpesvirus 3, 4, and 5 (porcine lymphotropic herpesvirus – PLHV-1, -2, and -3) are viruses that infect domestic and feral pigs.ObjectivesThis study examined the presence of PLHV DNA in biological samples from free-living wild boars circulating in a Brazilian geographical region with a high density of commercial domestic pigs.MethodsLung samples of 50 free-living wild boars were collected by exotic wildlife controller agents between 2017 and 2019 in the state of Paraná, southern Brazil. Lung and spleen fragments were obtained from six fetuses collected by hysterectomy post mortem from a pregnant sow. A polymerase chain reaction (PCR) assay using consensus primers (pan-herpesviruses) was performed to detect PLHV DNA. The samples showing positive results for PLHV DNA were submitted to single-round PCR assays with the specific primers for identifying PLHV-1 (213-S/215-As), PLHV-2 (208-S/212-As), and PLHV-3 (886s/886As). The specificity of the species-specific PCR products was assessed by nucleotide sequencing of the amplicons.ResultsForty-eight (96%) of the 50 lung samples analyzed were positive for PLHV by PCR using pan-herpesvirus primers. In 33 (68.75%) of the positive samples, at least two PLHV species were identified simultaneously. The DNA of PLHV-1, -2, and -3 was found in free-living wild boars of all ages, but not in the fetuses, even though they were from a sow that tested positive for all three viruses.ConclusionThese viruses are endemic to the population of feral pigs in the Brazilian region evaluated, as well as in domesticated pigs.

Evaluation of the Frequency of Swine Enzootic Pneumonia in one Farm Based on Slaughterhouse, Clinical and Laboratory Monitoring

Background: Swine enzootic pneumonia (SEP), caused by the bacterium Mycoplasma hyopneumoniae, is one of the main respiratory diseases of pigs. The purpose of this study was to evaluate the frequency of SEP in one farm with an independent production system, low level of technification and high losses in the meat packing plant, based on slaughterhouse, clinical and laboratory monitoring.Materials, Methods & Results: This study consisted of three monitoring steps, divided into three periods: P1, P2 and P3. In the first step of the study, slaughterhouse reports were analyzed and slaughtering was monitored at one meatpacking plant in Santa Maria, RS, Brazil. The second step of the study started by drawing up a profile of the farm that supplied the pigs to the aforementioned slaughterhouse, after which clinical monitoring was performed on three occasions. The third step of the study (laboratory monitoring) involved collecting lung fragments containing suspected SEP lesions from the meatpacking plant and subjecting them analysis. The average measured enzootic pneumonia (EP) in P3 was 38.54% (P < 0.05) higher than in P1 and 29.79 higher than in P2. The mean frequency of pulmonary emphysema (PE) in P3 was 59.13% (P < 0.05) higher than in P1 and 48.04% higher than in P2. The mean number of lung adhesions (LA) did not differ statistically (P > 0.05) between P1, P2 and P3. As for the mean frequency of pulmonary hepatization (PH), P3 was 48.80% higher (P < 0.05) than P1 and 41.78% higher than P2. With regard to the mean frequency of craniodorsal lung lesions (CLL), P3 was 48.26% higher (P < 0.05) than P1 and 40.77% higher than P2. The mean frequency of disseminated lesions (DL) and the pneumonia severity index (PSI) showed no statistical difference (P > 0.05) between the three evaluated periods. On the other hand, in the finishing period (FP), the frequency of coughing and sneezing was 37.56% (P < 0.05) higher in P1 than in P3. The frequency of coughing and sneezing evaluated in the nursery phase (CF) did not differ (P > 0.05) between periods. The samples sent to the laboratory MicroVet tested positive for M. hyopneumoniae, Influenza virus and Pasteurella multocida capsular type A. In the samples sent to the laboratory LABAC, the mean PCRmo (PCR for identification of mollicutes) did not differ (P > 0.05) in the analyzed periods, although the presence of DNA from bacteria of the class Mollicutes was confirmed in the fragments. In addition, the mean results of PCRmy (PCR for identification of M. hyopneumoniae) also did not differ (P > 0.05), although they revealed positivity for M. hyopneumoniae. Discussion: In relationship to the mean frequency of PE in each analyzed period, our study revealed an increase in the number of pulmonary lesions at slaughter during the periods under analysis. This finding is in agreement with those of other authors, who explain that pneumonias is one of the main health problems for swine producers. Some lesions are identified more frequently in routine inspections in meatpacking plants, including pulmonary emphysema, lung adhesions and pneumonia caused by M. hyopneumoniae. In our study, we found that the clinical signs of respiratory distress decline between the periods under evaluation, but during slaughter, the lesions suggestive of SEP continued to show an upward trend, mainly because they were chronic. Other authors state that several control measures should be adopted to ensure good zootechnical indices, including vaccination, strategic medication and the reduction of risk factors, thereby minimizing the level of M. hyopneumoniae infection in the herd.

Biological Evaluation of Azetidine-2-One Derivatives of Ferulic Acid as Promising Anti-Inflammatory Agents

The purpose of this study was to evaluate the in vivo biological potential of new azetidine-2-one derivatives of ferulic acid (6a–f). First, the in vivo acute toxicity of azetidine-2-one derivatives of ferulic acid on Swiss white mice was investigated and, based on the obtained results, it can be stated that the studied derivatives belong to compounds with moderate toxicity. The in vivo anti-inflammatory potential of these derivatives was determined in a model of acute inflammation induced by carrageenan in rats and in a chronic inflammation model induced in rats using the granuloma test. In the acute inflammation model, all the studied compounds had a maximum anti-inflammatory effect 24 h after administration, which suggests that these compounds may be classified, from a pharmacokinetic point of view, in the category of long-acting compounds. The most active compound in the series was found to be compound 6b. In the case of the chronic inflammation model, it was observed that the studied compounds (6a–f) reduced the formation of granulation tissue compared to the control group, having an intense effect of inhibiting the proliferative component. The most important inhibitory effect of inhibiting the proliferative component was recorded for compound 6b. Additionally, the investigation of liver function was performed by determining the serum levels of liver enzymes aspartate transaminase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH) and bilirubin (total and direct). The results showed that, in the series of azetidin-2-one derivatives, the liver enzymes concentration values were close to those recorded for the reference anti-inflammatories (diclofenac sodium and indomethacin) and slightly higher compared to the values for the healthy control group. At the end of the experiment, the animals were euthanized and fragments of liver, lung, and kidney tissue were taken from all groups in the study. These were processed for histopathological examination, and we noticed no major changes in the groups treated with the azetidine 2-one derivatives of ferulic acid compared to the healthy groups.

Glässer’s disease in swine from Northeastern Brazil

ABSTRACT: Glässer’s disease is an important infectious disorder of swine caused by Haemophilus parasuis. Although well recognized in most regions of Brazil, outbreaks of Glässer’s disease have not been described in Northeastern region. For this reason, three municipalities of the Pernambuco State were visited in order to identify histories of high mortality in growing and finishing pigs. The main clinical signs consisted of dry cough, apathy, fever, anorexia, paresis, muscle tremors, motor incoordination, seizures leading to high mortality rates. Nine pigs were necropsied, and fragments of the nervous system, organs of the abdominal and thoracic cavities were collected for histological analysis. In addition, lung and brain fragments were used for DNA extraction and molecular testing by real-time Polymerase Chain Reaction (PCR). Grossly, the main lesions consisted of petechial hemorrhages or ecchymosis on the skin of the face, abdomen, forelimbs, and hind limbs. The main severe lesions consisted of hydropericardium, hemopericardium, fibrinous pericarditis and pleuropneumonia. Microscopically, pericarditis, epicarditis and subepicardial myocarditis, followed by a moderate to severe multifocal pleuropneumonia, fibrinosuppurative and necrotizing were the most frequent lesions observed. Real-time PCR amplified H. parasuis infB gene in all samples analyzed, confirming the presence of this etiologic agent.

The lungworm, Halocercus brasiliensis (Nematoda: Pseudaliidae), from Guiana Dolphins Sotalia guianensis from Brazil with Pathological Findings.

The Guiana dolphin, Sotalia guianensis, is a small cetacean species found on the Atlantic coast of Central and South America from Honduras to Santa Catarina State, Brazil. The nematode Halocercus brasiliensis infects this cetacean, resulting in lung pathologies and death. The present study aimed to conduct a morphological and morphometric study of specimens of H. brasiliensis collected from S. guianensis from the coast of the state of Espírito Santo, Brazil. For this study, 7 dolphins were collected and examined for the presence of lung parasites. Nematodes were collected and processed for light microscopy, and lung fragments were processed for histological analysis. The nematodes were identified as H. brasiliensis according to their morphology and morphometry. The histopathological analysis revealed mineralization of the bronchiolar cartilage and inflammatory process. The parasitic infections by H. brasiliensis in S. guianensis can contribute to the debilitating status of these cetaceans, resulting in their stranding and accidental capture in fishing nets.

VIRUS-ASSOCIATED HEMOPHAGOCYTIC LYMPHOHISTIOCYTOSIS – THE SEVERE COURSE EXPRESSION IN SARS-COV-2 INFECTION?

The SARS-CoV-2 pandemic has caught the world completely unprepared. Sustained efforts are being made to understand the mechanisms by which this virus acts. One of the oldest and most efficient methods to achieve this is the autopsy, which becomes even more important as their number is limited. We present the case of an 88-year-old man, who died in a car, and was later autopsied at SCJML Constanta. The suspicion of SARS-CoV-2 infection was raised (the man had returned from the most affected area in Romania - Suceava) and the test was performed on a lung fragment taken during the autopsy. The presence of viral pneumonia with diffuse alveolar lesions was found. Lymphohistiocytic hemophagocytosis phenomenon was observed in paratracheal lymph nodes. In SARS-CoV-2 infection, there is the cytokine storm theory, which has multi-organ manifestation. It can occur in severe forms of infection with various viruses (Ebstein-Barr, SARS, MERS) and is the consequence of a hyperactive immune system, especially regarding macrophages. A morphological expression of this aberrant macrophage activation may be lymphohistiocytic hemophagocytosis.

Influence of different newborn piglets drying methods on the development of lesions in the respiratory tract

Delivery is one of the critical points of pig production, and the drying of newborns at this time is extremely important, since heat loss can reduce colostrum intake, cause starvation, diarrhea and crushing, with consequent increase mortality. Three main drying methods consist in: i) drying powder; ii) wood shavings; and iii) paper towels; being the first two composed of fine particles that could penetrate and cause damage to the respiratory tract. The objective of this research was to evaluate if the particle size of the material present in wood shavings and in the drying powder could lead to injuries to the respiratory tract of neonates. For this purpose, firstly, the wood shavings and the drying powder were subjected to particle size analysis. In addition, six litters were divided into three groups of 12 piglets each, and each group received a type of drying at birth. On the first, fourth and seventh days after birth, four piglets from each group were euthanized to collect diagnostic materials such as blood, for hematological evaluation, and fragments of lung and trachea for histopathological analysis. Hematological data showed no significant difference between groups, but within the wood shavings group there was a significant increase in lymphocytes on the seventh day, when compared with the first and fourth days. Histologically, the animals exposed to wood shaving and drying powder presented more severe histological findings compared to the paper towel group. Based on these results, the drying newborn piglets with wood shavings had higher potential to generate lesions in the lung tissue of newborn piglets.

Effect of piperlongumine during exposure to cigarette smoke reduces inflammation and lung injury

Chronic obstructive pulmonary disease (COPD) is related to smoking and anti-inflammatory therapy is indicated. Among the mediators with anti-inflammatory properties, we highlight piperlongumine (PL), an alkaloid/amide of Piper longum. Here we evaluated the PL administration on an experimental model of respiratory inflammation resulting from exposure to cigarette smoke. Male Balb/c mice were exposed to burning of 10 commercial cigarettes, 2x/day, for five weeks on specific equipment. PL efficacy was evaluated in control, exposed to smoke without treatment and PL treated (2.0 mg/kg, 3x/week) groups. Animals were weighed and plethysmographic analyses performed at the end of the exposure protocol. Inflammatory cells were evaluated in the bronchoalveolar lavage (BAL) and hemoglobin and glucose in the blood. Lung fragments were processed for histopathological studies and AnxA1, COX-2, NF-kB and neutrophil elastase expressions. Plethysmography revealed that PL maintained pulmonary frequency, volume and ventilation parameters similar to controls, with respiratory volume reduction compared to untreated animals. Final weight was reduced in both exposed groups. PL decreased hemoglobin concentration, attenuated the reduction of glucose levels and reduced influx of lymphocytes, neutrophils and macrophages in BAL. Histopathologically occured infiltration of inflammatory cells, increase of the interalveolar septa and intra-alveolar spaces in untreated animals. But, PL administration recovered lung tissues and, immunohistochemically, promoted increased expression of AnxA1 and reduction of COX-2, NF-kB and neutrophil elastase. Together the results indicate that PL attenuates systemic and pulmonary inflammatory changes, partially by modulating the expression the endogenous AnxA1, and may represent a promising therapy in preventing the inflammation induced by cigarette smoke.

Intercurrence of Paratuberculosis in Intradermal Tuberculin Test Reactive Cattle

Background: Bovine tuberculosis control programs are based on a standard diagnostic method, the intradermal test with purified protein derivatives, which is used to identify and eliminate diseased animals. Currently none of the tests available allow complete differentiation between infected and uninfected animals. The main limitations of the tests available are related to diagnostic sensitivity and specificity, which results in false positive reactions due to the existence of cross infections, and also false negative, inherent to the state of energy of some animals. The aim of this work was to study the intercurrence of paratuberculosis in tuberculosis reactive cattle by the comparative cervical test.Materials, Methods & Results: Three hundred and thirty four cattle were evaluated using the comparative cervical test (CCT) and serology for tuberculosis (TB) and paratuberculosis (PTB) ELISA IDEXX®. All of the animals testing positive by CCT were euthanized and necropsied. Fragments of lymph node, lung and intestine were collected and analyzed using histopathological techniques, with staining by Hematoxylin-Eosin (HE). Samples of lung and lymph nodes (retropharyngeal, submandibular, cervical and mediastinal) of the animals testing positive by CCT were evaluated using qPRC for M. bovis, and intestinal and mesenteric lymph nodes using PCR for PTB. Of the 334 cattle evaluated using the comparative cervical test, 16 were considered positive. No lesions suggestive of tuberculosis were found in the macroscopic inspection of the carcasses. The most evident anatomical and pathological finding was a thickening of intestinal mucosa, found in 12 of the 16 cattle submitted to necropsy. No microscopic lesions suggestive of TB were identified nor was the presence of M. bovis detected by qPCR. The main histopathological findings were observed in the small intestine and mesenteric lymph nodes and identified as enteritis, lymphangitis, lymphangiectasia and granulomatous lymphadenitis. In the intestine the changes are characterized by dilated and inflamed lymphatic vessels and intense inflammatory infiltrate on the mucosa and submucosa. Of the 334 serum samples evaluated, the M. bovis ELISA Antibody Test (IDEXX®) identified 17 positive animals. All the cattle considered positive by M. bovis ELISA were considered negative by CCT. In the samples from nine animals (9/16), DNA from M. avium subsp. paratuberculosis (MAP) was identified and in twelve carcasses (12/16) lesions characteristic of PTB were found, which were subsequently confirmed by histopathological techniques. In another nine animals of the herd anti-MAP antibodies were detected. None of those that tested positive by PTB ELISA were reactive by CCT.Discussion: Animals considered positive by TB ELISA that were not positive in the intradermal test does not mischaracterize the clinical picture of the disease. Considering the inverse relationship between cell-mediated and humoral responses to M. bovis, the intradermal test and the serological tests are designed to measure different immunological responses, which develop during different stages of infection. The progress of the cellular immunological response to humoral immunity occurs in the most advanced stages of tuberculosis. Of the 16 cattle considered positive by CCT, 12 animals presented macroscopic and histological lesions suggestive of PTB and DNA from MAP was detected in nine. Although it is the official test for the control of TB in different countries, the intradermal test with PPD has presented limitations, primarily related to specificity. M. avium subsp. Paratuberculosis is considered the main cause of false positive reactions in the intradermal test. The PPD bacterial extract is a complex mixture of proteins, lipids, sugars and nucleic acids, and many of these components are also shared by numerous species of mycobacteria (tuberculous or not).

Avaliação de duas técnicas de extração de DNA na detecção de Mycoplasma hyopneumoniae por PCR em amostras de pulmões de suínos com e sem lesão de pneumonia

Swine Enzootic Pneumonia (PES) has as the primary etiologic agent the Mycoplasma hyopneumoniae. The diagnosis of PES involves clinical manifestations on farms, macroscopic lesions visualized in the slaughterhouse and laboratory techniques, such as isolation by culture, histopathology, immunohistochemistry, ELISA and other serological tests and PCR. This work aimed to establish the comparison between PCR results and typical macroscopic findings in lungs with PES and to evaluate the efficacy of PCR results with extraction by phenol-chloroform and commercial kit in fragments of swine lungs. The efficacy of PCR was evaluated by comparing the classic method of DNA extraction by phenol-chloroform and by commercial Kit. Fragments of lungs of 18 pigs with characteristic lesions of PES and 17 without lesions were subjected to DNA extraction and posterior PCR. The pulmonary lesions with PES were PCR-positive in 94.44%, compared to 76.47% of the non-PES lungs. Kappa concordance between macroscopic and PCR was 18%, which was not significant (P > 0.05). Of the 35 lung fragments processed by phenol-chloroform, 85.71% were positive by PCR, compared to 28.57% by the commercial kit. Kappa agreement between DNA extraction methods was 57%, which was statistically significant (p <0.05), but Odds ratio was 15 (CI 4.52 to 49.68), which means that the probability of PCR positivity by the phenol-chloroform method was 15 times higher than that by the commercial kit. Macroscopy and PCR results were discordant, with phenol-chloroform extraction being the best between both procedures.

Immunohistochemical evaluation of surfactant-associated protein in fibrosis-cavernous pulmonary tuberculosis

Background. Despite the progress and development of scientific directions in various fields of medicine, the problem of tuberculosis and its morphological manifestations remains relevant and is not fully disclosed due to complex pathogenesis, the presence of various clinical forms, therapeutic pathomorphosis, torpid to therapy, the presence of relapses. It is known that a surfactant system occupies a special place in the system of local lung protection.Aim. To study the condition of surfactant-assotiated protein A in the foci of specific destruction and in the surrounding intact lung tissue to assess its functional status, degree of respiratory failure and possible dissemination of tuberculous inflammation.Material and Metods. An analysis of 163 lung fragments of the dead or operated on for cavernous pulmonary tuberculosis with active bacterial excretion of 89 fragments and with clinical abacilation – 74 was carried out. Results A morphological study revealed stereotypical dynamic depression of surfactant-associated protein A in all the samples studied, both in the areas of cavernous destruction and pericavernouse zone, and in intact lung tissue. The maximum intensivity of the immunohistochemical expression of this surfactant protein was recorded in the alveolar macrophages, which indicated intensive recycling and utilization of the components of the surfactant.Conclusion. Minimizing the production of surfactant components and its active utilization in intact lung tissue leads to a collapse of the alveoli with subsequent progression of respiratory failure.

Electron Microscopic Characteristics of The Blood-Air Barrier Interstitium in Fibrous Cavernous Pulmonary Tuberculosis in Comparison with Chronic Nonspecifi c Lung Diseases

Aim. To describe the ultrastructural characteristics of the blood-air barrier (BAB) interstitium in fi brous cavernous pulmonary tuberculosis (FCT) in comparison with chronic nonspecifi c lung diseases (CNSLD).Materials and methods. The fragments of the pericavernal zone and lung tissue were taken for the study at the resection border from the dead or operated for CNSLD persons (n = 163), and the perifocal and boundary zone of lung tissue. 116 CNSLD patients were divided into 3 subgroups: 1) chronic lung abscess (n = 42); 2) bronchiectasis (n = 44); 3) lung cyst (n = 30). The lung fragments of 30 patients who died from pathology not associated with lung diseases (myocardial infarction, acute cerebrovascular accident) were used as a control group to compare the morphological parameters. The criteria for inclusion of patients in the study: age from 18 to 65 years, negative clinical and laboratory data on the presence of comorbid pathology (viral hepatitis B, C and HIV). For TEM, lung fragments 1×1×1 mm in size were cut out and fi xed in a 2.5% glutaraldehyde solution in phosphate buffer (pH = 7.2–7.4) and washed in 0.1 M phosphate buffer (pH = 7.4), followed by dehydration in alcohols of an ascending concentration and placing in a mixture of Epon and Araldite resins according to the scheme. Ultrathin sections were made with Reynolds staining. Viewing and photographing preparations was carried out on a PEM-100 transmission electron microscope (Ukraine) (magnifi cation range from ×1000 to ×30 000). Results. It was established that changes in BAB components in all groups had similar features in the form of severe interstitial fi brosis, the signs of endothelial cell degeneration and destruction of varying degrees of severity, as well as the heterogeneity of the endothelial and epithelial basement membranes.Conclusion. Ultrastructural changes in the BAB components of the removed lung part in patients with FCT and chronic nonspecifi c lung diseases are characterised by a polymorphism with prevailing dystrophic and destructive changes in the perifocal zone of infl ammation, and compensatory-adaptive processes on the peripheral, especially at the resection border.

Morphofunctional changes in the heart and lung tissues of dogs for isoniazid poisoning

The article presents the results of macroscopic, histological and histochemical studies of the heart and lungs of dogs for isoniazid poisoning. A pathoanatomical section of 19 carcasses of dogs of different ages and breeds was performed; heart and lung fragments were selected for morphological examination. Heart and lung samples were fixed in 10 % aqueous formalin neutral solution, Carnua and Buen fluids. Dehydration was carried out in ethanol of increasing concentration and embedded in paraffin. The sections were made on a MC-2 microtome 7 μm thick, stained with hematoxylin and eosin, using the McManus and Brasche technique. Made histopreparations were examined under a Leica DM 2500 light microscope (Switzerland), photo-fixation was performed with a Leica DFC450C using Leica Application Suite Version 4.4 software. The results of morphohistochemical studies have shown that in acute poisoning of dogs with isoniazid in the heart prevailed acute disorders of the vascular circulation, manifested by a sharp expansion and destruction of the walls of the venous vessels, plasmorrhagia, microtromb of the capillaries and spasm of the arterioles, which led to the development of swelling, caused acute decrease of protein synthesation function of muscle fibers and dystrophic-necrobiotic changes of cardiomyocytes. It was noted, in the lungs of dogs on the background of acute congestive hyperemia, diapedesis of erythrocytes and accumulation of transudate in the lumen of the alveoli, foci of emphysema and bronchiectasis with rupture of the epithelial layer of the mucous membrane, elastic skeleton and muscular tubules of bronchial tubules that indicated the violation of gas exchange and the development of anoxemia. It is known that spontaneous isoniazid poisoning in dogs leads to an acute course and often fatal consequences. In the comprehensive study of the effects of chemicals on the body, and in particular isoniazid on the body of dogs, toxicological and pathomorphological studies are of great importance. The use of morphological and histochemical methods will reveal some of the mechanisms of intoxication.

Morphofunctional Transformations in the Lungs of Rats Under the Long-Term Exposure to Sodium Tetraborate

The aim of the study was to identify reactive and adaptive changes in the lungs of rats under the longterm exposure to sodium tetraborate.Material and methods. The study included male rats which were administered sodium tetraborate in dosage of 1/30 LD50, intraperitoneally, daily. The study samples (lung fragments) were selected for histological examination in 7, 14, 21 and 30 days from the beginning of the experiment.Results. Long-term exposure to sodium tetraborate resulted in a complex of destructive changes in the air-conducting and respiratory parts of the lung. Leukocyte infiltration in the connective tissue and epithelium of the bronchial and alveoli wall, focal destruction of the bronchial epithelium and alveoli, growth of the connective tissue in the organ interstitial were observed with underlying edema, stasis of blood corpuscles in capillaries, focal destruction of the capillary wall. The study revealed alveolocyte wall thickening and growth and sclerosing of the connective tissue in the interalveolar spaces; this appears to be the morphological equivalent of the increased thickness of the aero-hematic barrier and deterioration of the gas exchange in the alveoli. The increased proportion of the bronchi-associated lymphoid tissue mainly presented by the lymphoid tissue of the diffuse character and less rarely by the lymphoid follicles was registered in the wall of the medium bronchi.Conclusion. The results have proven the negative impact of sodium tetraborate on lung structures and demonstrated the adaptive capacity of the lungs, their ability to maintain the necessary structural-functional characteristics under the extreme destabilizing factors effect.

High prevalence of porcine circovirus 2, porcine parvovirus, and pathogenic leptospires in mummified swine fetuses in Southern Brazil

ABSTRACT: Modern swine production employs a high degree of technology and is organized in various production stages, in which reproduction is one of the most significant. However, reproductive losses associated to fetal death are still high. Fetal losses, including fetal mummification, may occur at a rate below 1.5% in a sound herd. The causes of fetal death can be very diverse, and include various infectious agents. Therefore, investigating the causes of fetal losses is important in the decision-making process related to the control and prophylaxis of the herd. The goal of this study was to carry out a molecular survey, analyzing porcine circovirus 2 (PCV2), porcine parvovirus (PPV), and pathogenic Leptospira as potential agents related to fetal death in swine farms in Southern Brazil. We collected samples from three farms with a mummified index ≥ 2.5%. Fragments of brain, lung, kidney, liver, and heart were sampled for PCR analysis. Out of 100 samples examined, 87, 68, and 22 were positive for PCV2, PPV, and Leptospira spp., respectively. Moreover, we also identified coinfections with two and three pathogens in the same sample. Our findings contribute to the spreading of scientific knowledge related to infectious causes (PPV, PCV2, and pathogenic leptospires) of fetal losses in swine, as evidenced by the high frequencies of the investigated agents.

Pathological and molecular characterization of avian malaria in captive Magellanic penguins (Spheniscus magellanicus) in South America.

Avian malaria is a mosquito-borne disease that affects multiple avian species and is caused by protozoans of the genus Plasmodium. An avian malaria infection caused by Plasmodium sp. in Magellanic penguins (Spheniscus magellanicus) with high mortality is described in a zoo in Southern Brazil. Clinically, three birds presented signs of inappetence, anorexia, pale mucosa, dyspnea, and opisthotonus, with death in a clinical course of 5-8h. At the necropsy, all birds exhibited pale mucosa, marked splenomegaly and hepatomegaly, in addition to moderate leptomeningeal blood vessels ingurgitation in the brain. Microscopically, multiple exoerythrocytic meronts were observed in the cytoplasm of endothelial cells in the spleen, liver, heart, lungs, brain, kidneys, and pancreas. The spleen had a multifocal perivascular inflammatory infiltrate of lymphocytes, plasma cells, and macrophages, which also exhibited hemosiderosis and erythrophagocytosis. The liver had a multifocal periportal inflammatory infiltrate of lymphocytes, macrophages, and plasma cells, in addition to marked hemosiderosis in the hepatic sinusoids. Fragments of spleen, liver, brain, skeletal muscle, and lung were tested by the polymerase chain reaction technique for the detection of a fragment of the cytochrome B gene from haemosporidians, which resulted positive for Plasmodium spp. After sequencing, the samples were phylogenetically associated to Plasmodium sp. detected in Turdus albicollis (KU562808) in Brazil and matched to the lineage TURALB01 previously detected in T. albicollis. Avian malaria infections caused by Plasmodium sp. of lineage TURALB01 may occur in S. magellanicus with high mortality, and, thus, it is essential to detect and characterize the agent involved to obtain the differential diagnosis of the condition.

Linfossarcoma de células B multicêntrico em uma ovelha

Background : : : Lymphosarcoma is a malignant tumor from tumoral cells similar to lymphocytes and its spontaneous development in small ruminants is unusual in Brazil. In cattle, the lymphosarcomas are often associated with infection by bovine leukemia virus (BLV), although sheep are considered as a very susceptible specie to BLV. Spontaneously lymphosarcoma is rare and in most cases it is observed in apparently healthy animals in slaughterhouses. There is no report of this tumor in sheep in southern Brazil. We describe a case of spontaneous multicentric B-cells lymphosarcoma in an ewe. Case : In June 2007, a 4-years-old mixed breed ewe was necropsied after clinical signs as anorexia, weight loss and death in few weeks. This ewe was purchased in September 2006 with another 44 sheep, most females of the same age. Dairy cattle were also raised together with sheep at the same farm. Fragments of liver, lung, spleen, kidneys, brain and parotid lymph node were sent to the Veterinary Pathology Laboratory - Universidade Federal de Santa Maria, Brazil, for histopathological evaluation. Multiply white to yellow nodules, with 0.5 to 2 cm of diameter, were observed in the liver, kidneys, and the parotid lymph node. The nodules were soft and some coalesced and infiltrating the hepatic parenchyma. The kidneys nodules were similar to those of the liver but prominent in the capsule. They were observed infiltrating more intensely the cortical region. The parotid lymph node was 6 x 4 x 3.5 cm size, whitish, multilobulated and the cut surface was totally obliterated by the tumor mass. Histologically, the neoplasm showed intense proliferation of neoplastic lymphoid cells arranged in sheets and a poor fibrovascular stroma, with round small and hyperchromatic nuclei, and scarce eosinophilic cytoplasm compressing the adjacent parenchyma. Immunohistochemistrily, neoplastic cells were strongly positive for anti-CD79a antibody and negative for anti-CD3 antibody, indicating neoplastic cells derived from B lymphocytes. Discussion : The diagnostic of multicentric B-cell lymphosarcoma was made based on gross, histological, and immunohistochemical findings. Spontaneous development of this tumor is uncommon in sheep in Brazil. The multicentric presentation seen in this case is the most common form of lymphosarcoma in sheep, although the thymic and cutaneous forms also occur. Sheep affected by multicentric lymphosarcomas may have symmetrical lymphadenopathy, not observed in this case, and involvement of various organs including liver, kidney, spleen, intestine, skeletal muscle and heart. However, under experimental conditions, these tumors are easily induced after inoculation of bovine retrovirus (BLV) in sheep, by its high sensitivity. Possibly, the spontaneous transmission of BLV from cattle developing the disease in sheep may occur in Brazil as well as in other countries. Epidemiological factors of the disease are not established yet. Despite this, some correlation must exist for the low occurrence of this disease in sheep in properties of southern Brazil, where cattle and sheep are commonly raised together, and the frequency of lymphosarcoma in sheep is low. In this case, the association of BLV with development of the neoplasm could not be confirmed. This is the first report of multicentric B-cell lymphosarcoma in sheep diagnosed in southern Brazil.

Congenital cystic adenomatoid malformation - diagnostic and therapeutic procedure: 8-year experience of one medical centre.

IntroductionCongenital cystic adenomatoid malformation (CCAM) is a rare anomaly. The mechanisms and the time at which the abnormality develops are still unclear. The malformation is characterized by the presence of single large or multiple but smaller cysts.AimTo present the experience of our medical centre, the Polish Mother’s Memorial Hospital – Research Institute.Material and methodsWe analysed the medical records of 32 neonates hospitalized in 2008–2017 at the Department of Paediatric Surgery and Urology ICZMP due to pre- or postnatally diagnosed congenital cystic adenomatoid malformation.ResultsIn 2008–2017 32 children were hospitalized at our department due to prenatally diagnosed CCAM. An intrauterine procedure of thoracoamniotic shunting was performed in 13 fetuses. All newborns had chest computed tomography. Surgery had been performed in 21 children. Histopathological analysis of specimens prepared from the resected lung tissue confirmed type 1 CCAM in 4 cases, type 2 CCAM in 11 cases and type 3 CCAM in 1 case. Four patients were diagnosed with a bronchogenic cyst and one with extralobar pulmonary sequestration.ConclusionsPostnatal management of CCAM depends on the clinical status of the newborn. Newborns without clinical symptoms require radiological diagnostics to confirm the presence of malformation. Surgical treatment is recommended before the age of 10 months. Resection of the malformed lung fragment in patients with CCAM is the first-line treatment. It should be emphasized that despite the tremendous advances in diagnostic imaging, the final diagnosis of CCAM is made based on histopathological findings.

Diatoms in lung tissue: first investigation in Brazil in proving death by drowning

Diatoms are microalgae widely distributed in inland and marine waters and during drowning they are inhaled together with water, passing through the bloodstream to various internal organs of a victim. Our investigation aimed to evaluate the diatoms extracted from the lung tissues of an individual with drowning diagnosis and compare them to those found in the environment, in order to prove the cause of death and the drowning site. A left lung fragment of 17.9 grams removed from the victim was oxidized and prepared for diatoms identification and quantification using inverted microscope. A total of 133 diatoms valves was found, being 85% belong to pennate forms and 15% to centric forms. Pinnularia, a bentonic genus, had greatest number of valves, followed by Fragilaria, Actinocyclus, Cocconeis and Encyonema. These results revealed that the victim drowned at the bottom of the lake, inhaling both water and sediments with diatoms. This is the first investigation of this nature carried out in Brazil.