Polyvalent Bacterial Lysate with Potential Use to Treatment and Control of Recurrent Urinary Tract Infections.

Overuse of antimicrobials has greatly contributed to the increase in the emergence of multidrug-resistant bacteria, a situation that hinders the control and treatment of infectious diseases. This is the case with urinary tract infections (UTIs), which represent a substantial percentage of worldwide public health problems, thus the need to look for alternatives for their control and treatment. Previous studies have shown the usefulness of autologous bacterial lysates as an alternative for the treatment and control of UTIs. However, a limitation is the high cost of producing individual immunogens. At the same time, an important aspect of vaccines is their immunogenic amplitude, which is the reason why they must be constituted of diverse antigenic components. In the case of UTIs, the etiology of the disease is associated with different bacteria, and even Escherichia coli, the main causal agent of the disease, is made up of several antigenic variants. In this work, we present results on the study of a bacterial lysate composed of 10 serotypes of Escherichia coli and by Klebsiella pneumoniae, Klebsiella aerogenes, Enterococcus faecalis, Proteus mirabilis, Citrobacter freundii, and Staphylococcus haemolyticus. The safety of the compound was tested on cells in culture and in an animal model, and its immunogenic capacity by analysing in vitro human and murine macrophages (cell line J774 A1). The results show that the polyvalent lysate did not cause damage to the cells in culture or alterations in the animal model used. The immunostimulatory activity assay showed that it activates the secretion of TNF-α and IL-6 in human macrophages and TNF-α in murine cells. The obtained results suggest that the polyvalent lysate evaluated can be an alternative for the treatment and control of chronic urinary tract infections, which will reduce the use of antimicrobials.

Gastrointestinal helminths of waterfowl (Anatidae: Anatinae) in the Lerma marshes of central Mexico: Some pathological aspects.

Gastrointestinal helminths of migratory waterfowl can cause several lesions that may affect the health and even the survival of their hosts. As part of an ongoing project that aims to inventory the helminth species that infect this group of birds, as well as the histopathologic lesion they cause, a total of 200 digestive tracts of nine species of anatid birds (Spatula discors, S. cyanoptera, S. clypeata, Mareca strepera, M. americana, Anas crecca, A. acuta, A. platyrhynchos diazi, and Oxyura jamaicensis) were collected in the Atarasquillo marsh of Lerma, in the State of Mexico. The present work had a twofold goal: to determine the taxonomic identity of the helminth species present in waterfowl as well as their infection levels and to describe the lesions caused by gastrointestinal parasites in migratory and resident anatids in Atarasquillo marsh, State of Mexico. The specimens were examined using routine helminthological and histopathological techniques. A total of 23 helminths were identified: six trematodes (Zygocotyle lunata, Notocotylus triserialis, Notocotylus seineti, Psilochasmus oxyurus, Australapatemon burti and Cotylurus magniacetabulus), four cestodes (Cloacotaenia megalops, Fimbriaria fasciolaris, Gatrotaenia cygni and Diorchis sp.), eleven nematodes (Echinuria uncinata, Tetrameres sp., Tetrameres fissispina, Hystrichis sp., Streptocara sp., Amidostomum sp., Epomidiostomum uncinatum, Capillaria sp., Capillaria contorta, Ascaridia sp. and Heterakis sp.) and two acanthocephalans (Pseudocorynosoma constrictum and Filicollis sp.). At the proventriculus level, Echinuria uncinata caused the most severe lesions; in the gizzard, Amidostomum sp. and E. uncinatum produced hemorrhages and necrosis. Finally, the main lesions found in the intestine (necrosis, heterophilic granulomas and the formation of lumps on the serosa) were caused by the implantation of the acanthocephalans' proboscis. Eleven species of gastrointestinal helminths found are new records for the Lerma marshes, as well as the first record in Mexico of the nematode Capillaria contorta and the acanthocephalan Filicollis sp.

Detection and Molecular Identification of Sarcocystis rileyi (Apicomplexa: Sarcocystidae) From a Northern Shoveler (Anas clypeata) in Mexico.

We detected macroscopic Sarcocystis cysts in a Northern Shoveler ( Anas clypeata ) in the Lerma Marshes, State of Mexico, Mexico in February 2014. The 5.0×2.0-mm macrocysts in the breast muscle of the duck were ovoid and yellow. Using an optical microscope, we saw parasitic forms of a Sarcocystis sp. among muscular fibers; the cysts measured 3.5×1.1 mm. The external wall had a smooth surface and the internal wall had a spongy texture. We identified the macrocysts as Sarcocystis rileyi according to sequences of the 18S rRNA gene, 28S rRNA gene, and ITS-1 region. Sarcocystosis should be considered in similar assessments in wild waterfowl in Mexico. Awareness of S. rileyi among anatids in the Lerma Marshes will contribute to more-effective conservation and management actions.

Toxoplasmosis en una colonia de monos ardilla (Saimiri sciureus) en cautiverio en Cuernavaca, Morelos, México / Toxoplasmosis outbreak in a private collection of squirrel monkeys (Saimiri sciureus) in Cuernavaca, Morelos Mexico

Toxoplasmosis is a disease of worldwide distribution that parasites the intestinal tract of wild and domestic cats. However, in New World monkeys it is considered an acute and fatal illness. The reason for its high susceptibiliy in this particular species is still unknown, but it has been theorized that their habitat and living conditions, in high trees, might be an important factor. Necropsy was performed on 5 squirrel monkeys (Saimiri sciureus), three males (adult) and two females (one of them gravid), with a clinical history of eye redenning and presence of bloody serous foam in the nostrils. Gross examination of the lung showed non collapsed lobes, dark red parenchyma and irregular pale pink nodular areas. All five animals showed apparent hepatomegaly. Histological findings revealed an interstitial pneumonia with a prominent thickening of alveolar septae, and numerous foamy macrophages with intracytoplasmic basophilic structures 20- 25 urn long, covered by a refringent wall and housing PAS positive basophilic structures that were aproximately 8 urn long (bradyzoites), which were identified as protozoal cysts. The liver showed multiple foci of periportal inflammatory cells and numerous parasitic cysts that were also observed in spleen and brain tissue. Inmunohistochemical technique was performed on selected samples of brain, liver and lung tissue, which resulted positive for Toxoplasma gondii using antibodies of hyperimmune antiserum-caprine 6244 origin, in a 1:300 dilution. Samples of the same tissues were processed for transmission electron microscopy. Hepatocytes of the liver showed multiple intracytoplasmic vacuoles with parasitic structures consistent with tachyzoites, which were 3.6 μm long and measured 2.3 μm in diameter. They possesed a slightly wavy electrodense plasmatic membrane that exhibited a conoid in its anterior end, along with several electrodense micronemes. Ultraestructural features like the conoid and the location of micronemes allowed us to identify the parasite as Toxoplasma gondii. Clinical history, necropsy findings, microscopic examination, inmunohistochemical analysis and electron microscopy of all five squirrel monkeys were consistent with a diagnosis of systemic toxoplasmosis. This report shows the high suceptibiliity of New World monkeys to become infected with this parasite. Therefore, toxoplasmosis should be considered among the differential diagnosis for diseases with non specific clinical signs in this particular species.

La toxoplasmosis es una enfermedad de distribución mundial provocada por el protozoario Toxoplasma gondii cuyos huéspedes definitivos son los felinos y una gran diversidad de especies animales son los huéspedes intermediarios. En primates del Nuevo Mundo se caracteriza por ser una enfermedad aguda y de curso fatal. La razón de la alta susceptibilidad en estos animales hasta la fecha se desconoce y generalmente se ha asociado con sus hábitos arborícolas. Se realizó la necropsia de cinco monos ardilla (Saimiri sciureus), tres machos (adultos) y dos hembras (una de ellas gestante), con historia clínica de enrojecimiento de los ojos y salida de espuma serosanguinolenta por la nariz. Macroscópicamente, los cinco animales presentaron los lóbulos pulmonares no colapsados y el parénquima rojo oscuro con áreas rosa claro entremezclado que en ocasiones tenían aspecto nodular. El hígado de los cinco animales se apreció aumentado de tamaño y pálido en su totalidad. Microscópicamente se observó neumonía intersticial caracterizada por el engrosamiento de los septos alveolares y la presencia de numerosos macrófagos espumosos cuyo citoplasma contenía estructuras elongadas, basofílicas de 20-25 μm de diámetro con pared refringente que a su vez albergaban estructuras basofílicas de 8 um (bradizoítos) PAS positivas consistentes con quistes de protozoarios. El hígado presentó focos de infiltrado inflamatorio periportal, así como numerosos quistes de protozoarios, que también se apreciaron en bazo y cerebro. Se realizó inmunohistoquímica de secciones histológicas de pulmón, hígado y cerebro, las cuales fueron positivas para Toxoplasma gondii con el anticuerpo Hyperimmune antiserum-caprine, a una dilución de 1:300. Asimismo, muestras de estos mismos tejidos fueron procesadas para microscopía electrónica de transmisión, en las que el hígado exhibía en su citoplasma vacuolas parasitarias con estructuras correspondientes a taquizoítos que medían de 3.6 μm de largo por 2.3 μm de diámetro, con una membrana plasmática electrodensa, ligeramente ondulada, que en ocasiones exhibían en su extremo anterior el conoide, y por debajo de éste, escasos micronemas electrodensos. Las características ultraestructurales como el conoide, y la ubicación de los micronemas, son criterios primordiales que permiten identificar al parásito como Toxoplasma gondii. La historia clínica, los hallazgos a la necropsia, el estudio histopatológico, la inmunohistoquímica y microscopía electrónica de los cinco monos ardilla fueron consistentes con un cuadro de toxoplasmosis sistémica. Este informe demuestra la alta susceptibilidad que poseen los primates del Nuevo Mundo al ser infectados por este protozoario, por lo que se debe considerar entre los diagnósticos diferenciales para enfermedades de curso clínico no específico en estos animales.

Presentación sistémica de Aspergillus spp con semiología neurológica en un Pastor Alemán: informe de un caso clínico / Systemic presentation of Aspergillus spp with neurological involvement in a German Shepherd: clinical case report

Infections of the central nervous system are uncommon in dogs. Pathogenic fungi such as Aspergillus, Cryptococcus spp, Blastomyces dermatitidis and Coccidioides immitis have been reported. An Aspergillus infection is mainly a respiratory event and very seldom it may become disseminated, the German shepherd seems to be the more prone to this type of aspergillosis specially if an immunological deficiency is present. A 1.5 year old female German Shepherd from Cancun, Quintana Roo, State, in the southeast of Mexico was presented to the small animal hospital, she showed tetraparesis, hiporeflex and deep retarded sensitivity in all four members, loss of sensitivity in the left side of the face, absence of pupil reflex and menace reflex as well as absence of muscular tone of the vulva and anal sphincter. The third day after presentation, she died. At necropsy polyencephalomalace, mielitis and diffuse leucomielitis piogranomalotosus were found. In the cytology of lymph node and other tissues aspergilli like hiphae were observed, this was confirmed by using the highly specific double diffusion test. Then, a PCR test was carried out with tissue fixed in 10% formaldehyde, but it failed probably due to destruction of the DNA.

La infección del sistema nervioso en el perro es poco común; posibles causas son Aspergillus spp, Cryptococcus neoformans, Blastomyces dermatidis y Coccidioides immitis. La infección por Aspergillus spp afecta al aparato respiratorio y en pocas ocasiones se presenta de manera diseminada. Sin embargo, en el Pastor Alemán hay mayor incidencia de éste, pues dicha infección se asocia con una deficiencia inmunológica. Se describe el caso de una perra de raza Pastor Alemán, de 1.5 años de edad, referida de Cancún, Quintana Roo, México, con tetraparesis, hiporreflexia y sensibilidad profunda retardada en los cuatro miembros, disminución de sensibilidad en el lado derecho de la cara y ausencia de reflejo pupilar y de amenaza en ojo derecho; además de ausencia de tono vulvar y de esfínter anal. A los tres días después del ingreso la perra falleció. A la necropsia se encontró polioencefalomalacia, mielitis y leucomielitis piogranulomatosa difusa. En la citología de linfonodo y en la histopatología de diferentes tejidos se identificaron hifas compatibles con Aspergillus spp. Con el fin de dar certeza al diagnóstico, se realizó prueba de doble inmunodifusión, que es altamente específica, la cual confirmó el diagnóstico de Aspergillus spp. Posteriormente se corrió PCR a partir de una muestra de tejido en formalina al 10%, para identificar la especie, en donde no se lograron resultados, probablemente debido a la destrucción del ADN por el formaldehído.

Mieloencefalitis protozoaria equina. Primer informe de una caso en México / Equine protozoal myeloencephalitis. A case report

Se solicita atención clínica para un caballo de raza Warm-Blood, 9 años de edad, que presentó: depresión, emaciación, incoordinación, disfagia; signos que se acentuaron durante siete días. Así mismo se observó: deshidratación, temperatura de 38.2 ºC, taquicardia, taquipnea, estupor, hipoalgesia facial, parálisis facial izquierda. Ceguera, pupila dilatada y estrabismo del ojo izquierdo, claudicación del miembro pelviano izquierdo. Se instauró tratamiento con sulfa-trimetroprim, dimetilsulfóxido y dexametasona. Al día 8 se presentó postración, marcada debilidad y movimientos de carrera. El reporte del líquido cefalorraquídeo indica elevación en la concentración de IgG (17.3 mg/dL), la prueba de Western blot y la reacción en cadena de la polimeraza resultaron positivas a mieloencefalitis protozoaria equina. El día 10 se decide eutanaciar al animal. A la necropsia se observó necrosis licuefactiva en lóbulo piriforme izquierdo, núcleo caudado izquierdo, rodilla del cuerpo calloso, pedúnculos cerebrales y médula espinal. El estudio histopatológico determinó en sección de cerebro medio y médula espinal cervical necrosis licuefactiva, abundantes células inflamatorias en leptomeninges y espacios perivasculares y estructuras parasitarias de forma rosetoide compatibles con protozoarios. Tales hallazgos establecen un diagnóstico morfológico de: meningoencefalitis parasitaria crónica multifocal grave. Los resultados del examen neurológico, evaluación de LCR e histopatología, sugieren 100 por ciento de seguridad de presencia de Sarcocystis neurona , agente etiológico de la mieloencefalitis protozoaria equina.