Effects of malathion on cellularity and sperm differentiation in testis and epididymis of adult rats / Efectos del malatión en la celularidad y diferenciación de espermatozoides en testículo y epidídimo de ratas adultas

Malathion is an organophosphorous insecticide, used worldwide for pest and disease control; however, it could also affect the reproductive patterns of several species. The aim of this study was to determine the effects of malathion in the cellularity and sperm differentiation in testis and epididymis of rats. Twenty adult male Sprague Dawley rats were divided into a malathion-treated group (n=10, dose of 170 mg/kg via subcutaneous injection for a period of 13 days) and control group (n=10, injected only with normal saline). After treatments, the rats were sacrificed by regulated euthanasia and assessed for sperm count in testis and epididymis and epididymal teratospermia degree. The results showed a significant decrease in body, testicular and epididymal weight in animals treated with malathion. Testicular sperm counts in treated rats exhibited a significant decrease in the number of sperm compared to controls (42.56x106 vs. 95.99x106), as well as in epididymis (77.55x106 vs. 106.54x106). Concerning the degree of teratospermia, a significant increase of abnormal sperm in the epididymis of treated rats versus controls (42.1% vs. 21%, respectively) was observed. We conclude that malathion has a cytotoxic effect in rats, significantly reducing the number of sperm produced by the seminiferous tubules and affecting their quality and number during the process of maturation and capacitation in their transit through the epididymis, thus increasing the level of teratospermia.

El malatión es un insecticida organofosforado, ampliamente usado en el control de plagas y pestes, sin embargo también puede afectar a los patrones reproductivos de las especies. El objetivo de este trabajo fue determinar los efectos de malatión en la celularidad y diferenciación de espermatozoides en testículo y en epidídimo de ratas. Veinte ratas macho adultas de la cepa Sprague Dawley, fueron divididos en grupo tratado con malatión (n=10) en dosis de 170 mg/kg de peso, inyección sub cutánea (s.c.), por un período de 13 días (duración del ciclo del epitelio seminífero) y grupo control (n=10), los cuales solo fueron inyectados con suero fisiológico. Finalizado el tratamiento las ratas fueron sacrificadas por eutanasia normada y se procedió a medir el recuento espermático en testículo y epidídimo y el grado de teratospermia en epidídimo. Los resultados obtenidos muestran una disminución significativa en el peso corporal, testicular y del epidídimo de ratas machos tratados con malatión. El recuento espermático en testículo de ratas tratadas, al compararlos con ratas controles, muestra una disminución significativa en el número de espermatozoides (42,56x106 / 95,99x106), igual comportamiento se observó en epidídimo (77,55 x106 / 106,54 x106). Al determinar el grado de teratospermia se observó un aumento significativo de espermatozoides anormales, en el epidídimo de las ratas tratadas versus los controles (42,1% y 21%, respectivamente). Se concluye que malatión tiene un efecto citotóxico en ratas, disminuyendo significativamente el número de espermatozoides producidos por los túbulos seminíferos y afectando la calidad y el número de ellos durante el proceso de maduración y capacitación, en su tránsito por el epidídimo, aumentando el nivel de teratospermia.

Malathion Affects Spermatogenic Proliferation in Mouse / Malathion Afecta la Proliferación Espermatogénica del Ratón

The restriction of the mechanisms of cell proliferation in murine seminiferous epithelium, in terms of induction of programmed cell death until recently has not been fully analyzed. The aim of this work was to assess the effect of Malathion (MP) on testicular morphology and function in mouse spermatogenesis. For the experiments, male albino mice of strain NMRI-IVIC, weighing between 30-40 g were used, and divided into control and experimental groups of 5 each. The animals of the experimental groups were injected with a single dose of MP: 241mg/kg weight (1/12 LD 50 ) resuspended in 0.9% saline, intraperitoneally. Animals were sacrificed at 8.3, 16.6 and 33.2 days post-injection (first, second and third spermatogenic cycles). Testicular samples were obtained for light microscopy (LM), transmission electron microscopy procedures, and to detect apoptosis and p53 antigen by immunohistochemical methods. Blood was collected to quantify testosterone and plasmatic cholinesterase activity. From 8.3 days, Sertoli cell vacuolization, karyolisis of pachytene spermatocytes and Leydig cells and a decreased in average of the diameter of seminiferous tubules was observed. No damage to inter-Sertoli cells junctions was detected. Percentage of seminiferous tubules showing germ cells apoptosis was increased from 8.3 days, plasmatic acetylcholinesterase activity was reduced in the group treated only 24 hours after administration of MP. Serum testosterone levels were low in treated animals at 16. 6 and 33.2 days. p53 was mostly expressed in pachytene spermatocytes from 8d. The findings of this study indicate that MP alters the testicular function affecting the DNA and interfering with spermatogenesis as well as steroidogenesis.

La restricción de los mecanismos de proliferación celular en epitelio seminífero murino, en términos de inducción de muerte celular programada hasta hace poco no había sido completamente analizada.El objetivo de este trabajo fue evaluar el efecto de malathion (MP) sobre la morfología y la función testicular del ratón.Ratones macho albinos de la cepa NMRI-IVIC, con pesos entre 30-40 g fueron utilizados, se dividieron en grupos control y experimental. Los grupos experimentales fueron inyectados por vía intraperitoneal con una dosis única deMP:241mg/kg de peso (1/12 DL50) resuspendido en 0,9% de solución salina.Los animales fueron sacrificados en el día 8,3, 16,6 y 33,2 después de la inyección (primer, segundo y tercer ciclos de la espermatogénesis).Se obtuvieron muestras de testículo para estudio en microscopía de luz (ML), microscopía electrónica de transmisión, para la detección de apoptosis y el antígeno p53 (proliferación celular), por métodos inmunohistoquímicos.Se recogió sangre para cuantificar la testosterona y la actividad plasmática de colinesterasa.Desde el día 8,3 día se observó vacuolización de células de Sertoli, cariolisis de espermatocitos en paquiteno y células de Leydig, y una disminución en el promedio del diámetro de los túbulos seminíferos. No se detectó daño en las uniones entre células de Sertoli. El porcentaje de túbulos seminíferos que mostraban células germinales en apoptosis se incrementó a los 8,3 días, laactividad de la acetilcolinesterasa plasmática se redujo en el grupo tratado sólo 24 horas después de la administración de MP.Los niveles séricos de testosterona disminuyeron en los animales tratados a los 16,6 y 33,2 días.P53 se expresó sobre todo en los espermatocitos en paquiteno desde los 8,3 días.Los resultados de este estudio indican que MP altera la función testicular, afecta al ADN e interfiere con la espermatogénesis, así como con la esteroidogénesis.

Protective effect of melatonin on damage in the sperm parameters of mice exposed to diazinon / Efecto protector de la melatonina sobre el daño en los parámetros espermáticos de ratones expuestos a diazinón

Since normal sperm parameters can be altered by organophosphorous pesticides, this study intended to determine if melatonin is able to prevent the damage on sperm quality after an acute exposure to diazinon. Adult male mice were injected intraperitoneally with melatonin, diazinon (1/3 or 2/3 LD50) or both, and sperm parameters were evaluated on days 1 or 32 post injection. Groups treated with diazinon showed elevated lipid peroxidation levels on day 1 post treatment, while groups pretreated with melatonin before diazinon showed no difference compared to control. Sperm count showed a significant decrease in both diazinon-treated groups only on day 32 post injection; no differences were observed in groups pretreated with melatonin prior to diazinon compared to control. The percentage of abnormal sperm morphology increased in the diazinon-treated groups only on day 32 postinjection. The administration of melatonin prior to exposure to diazinon prevents the alteration of sperm parameters commonly caused by organophosphates, possibly due to its antioxidant properties.

Debido a que los parámetros normales de los espermatozoides pueden ser alterados por algunos contaminantes como los pesticidas organofosforados, este estudio pretende determinar si melatonina es capaz de prevenir o proteger del daño en la calidad espermática, después de una exposición aguda a diazinon. Ratones machos adultos fueron inyectados via intraperitoneal con diazinon 1/3 y 2/3 de la LD50 y otro grupo tratados con melatonina + 1/3 diazinon LD50 y melatonina + 2/3 LD50. Los parámetros espermáticos fueron evaluados al día 1 y al día 32 post tratamiento. Los grupos tratados con diazinon solo o conjugado con melatonina mostraron un incremento significativo en los niveles de lipoperoxidación en el tratamiento después de un día. Al día 32 no se observan diferencias significativas con el grupo control. El recuento espermático al día 1 no presenta diferencias entre los grupos tratados y el control. Sin embargo al día 32 los grupos tratados con diazinon solo, muestran una disminución significativa, solo el grupo de melatonina +1/3 diazinon, presenta valores similares al grupo control. La morfología espermática normal presenta una disminución significativa en grupos tratados con diazinon, pero un aumento significativo al día 32 en los grupos tratados con melatonina. Los mayores porcentajes de anormalidades se presentan en la cabeza y la cola de los espermatozoides. La administración de melatonina antes de la exposición al diazinon evita las alteraciones de los parámetros espermáticos, comúnmente causada por organofosforados, posiblemente debido a sus propiedades antioxidantes.

Epiblast embryo stem cells give origin to adult pluripotent cell populations: primordial germ cell, pericytic and haematopoyetic stem cells: a review / Las células madre del epiblasto dan origen a poblaciones de células pluripotentes adultas: células germinales primordiales, células madre pericíticas y hematopoyéticas: revisión

Adult stem cells are great promise to the future of regenerative therapy, and understanding of its embryonic origin permit the discrimination of stem cell sources. Embryonic stem cells derived from inner cell mass of blastocyst originate the primordial germ cells, and pericyte stem cell associated to vessels endothelium in yolk sac. Currently, it is being proposed that embryonic primordial germ cell could originate hematopoietic stem cells based on the detection of germ cell markers (SSEA-1/TEC-1, Oct-4 and Nanog) in stem cell harvested from fetal liver and bone marrow. However, different experimental evidence points at two separate differentiation routes toward primordial germ cells, and hematopoietic stem cell with the same embryonic origin. The expression of undifferentiated stem cell markers in umbilical cord and placental vessels, such CD34, CXCR4, c-kit and OCT4 demonstrates the intimate relation between pericyte stem cells, endothelium, haematopoiesis, and primordial germ cells, which all originate from embryonic stem cell from the inner cell mass epiblast.

Las células madre adultas son una gran promesa para el futuro de la terapia regenerativa, y la comprensión de su origen embrionario permite la discriminación de las fuentes de células madre. Las células madre embrionarias derivadas del macizo celular interno del blastocisto originan las células germinales primordiales, y células madre pericíticas asociadas al endotelio de los vasos del saco vitelino. En la actualidad, se propone que las células germinales primordiales embrionarias podrían originar a las células madre hematopoyéticas sobre la base de la detección de marcadores de células germinales (SSEA-1/TEC-1 oct-4 y Nanog) en células madre extraídas de hígado fetal y médula ósea. Sin embargo, diferentes evidencias experimentales apuntan hacia dos vías separadas de diferenciación en células germinales primordiales, y en células madre hematopoyéticas con el mismo origen embrionario. La expresión de marcadores de células madre no diferenciadas en el cordón umbilical y los vasos de la placenta, como CD34, CXCR4, c-kit y OcT4 demuestra la íntima relación entre las células madre pericíticas, el endotelio y las células germinales primordiales, las que se originan en células madre embrionarias a partir del epiblasto del macizo celular interno.

Cypermethrin effects on the adult mice seminal glands.

The insecticide cypermethrin acts upon the sodium channels. Their effects over animal health are not understood. Here, the effects of cypermethrin on the seminal glands (SGs) are studied (1/5 DL50 i.p.). Forty-five adult mice (CF1) were distributed in three groups: (1) untreated, (2) vehicle (oil), and (3) experimental (cypermethrin in oil). The animals were sacrificed at 1 and each 8.6 days. The SGs were processed for histology: Haematoxylin/P.A.S, Thyonin (0.6%) and Immunohistochemistry (Ki-67). In the SGs was quantified: the epithelium height, mastocytes, and cell proliferation. In the results, cypermethrin exerts an intense effect on epithelium height and cell proliferation. A net increase of both parameters was observed at 24 h (p0.05). However, the mastocytes increased drastically and progressively during the experimental period (p0.05). Then, the effects have acute manifestations, which would be responsible for the potential changes in the male's reproductive potentiality.

Melatonin prevents damage elicited by the organophosphorous pesticide diazinon on the mouse testis.

Organophosphates like O,O-diethyl O-2-isopropyl-6-methyl pyrimidinyl-4-g-1-phosphorothioate (diazinon) are pesticides used worldwide, which can affect both animals and man even after a single exposure. Whereas their toxicity is due to acetylcholinesterase inhibition, their secondary toxic effects have been related to free oxygen radicals. This study evaluates the effects of a single dose of diazinon and melatonin-a powerful antioxidant-on plasmatic acetylcholinesterase activity and testis histopathology in adult mice 1 and 32 days post-treatment. Diazinon diminished the plasma acetylcholinesterase activity on day 1 post-treatment, although testosterone levels remained unaffected. Morphometrical analysis showed a decrease in seminiferous epithelium height (days 1 and 32), whereas an increase in testicular superoxide dismutase (SOD) activity was detected (day 32). Melatonin pretreatment prevented every alteration induced by diazinon, except the diminution of acetylcholinesterase plasmatic activity. Testicular damage might be due to elevated concentrations of free oxygen radicals released upon diazinon exposure, inducing alterations in the DNA and promoting local apoptosis; however, antioxidant pretreatment with melatonin prevents or diminishes this damage.

Melatonin prevents damage elicited by the organophosphorous pesticide diazinon on mouse sperm DNA.

Toxic effects of pesticides are commonly associated with DNA damage. To evaluate the effect of the organophosphate diazinon on sperm DNA and to test whether melatonin could prevent this damage, male mice were intraperitoneally treated with melatonin, diazinon (1/3 or 2/3 LD50) or both; cauda epididymal spermatozoa were obtained on days 1 and 32 postinjection and tested for DNA alterations. On day 1, sperm from diazinon-treated mice showed augmented DNA breakages and reduced chromatin packaging, whilst DNA damage increased only in the diazinon 2/3 LD50 group. Micronucleus test of bone marrow cells demonstrated somatic cell chromosomal damage in both diazinon-treated groups. Pretreatment with melatonin before diazinon acute administration improved all parameters studied on day 1 pi. The organophosphorous pesticide diazinon is a dose-dependent testicular toxicant that alters the sperm DNA structure; melatonin is able to prevent this damage.

Histopathological and Histometrical Assessment of Boron Exposure Effects on Mouse Spermatogenesis / Evaluación Histopatológica e Histométrica del Efecto en la Espermatogénesis por Exposición a Boro

Boron is a chemical element widely used in many industrial activities. Exposure to it affects many organs in mammals, mainly reproductive male organs. This work evaluates boron effect in testis. For this purpose, 12mg Boron/L of drinking water, equivalent to Arica tap water, was given for 42 days to 85 days old CFl male mice (experimental group). Another group (control group) drunk tap water of Santiago (0.6mg Boron/L) was used. Testicular histopathology and morphometric analysis was done. These studies showed that Borum induces alterations such as epithelial vacuolization, blockage of the tubular lumen and atrophy. Morphometrical data showed that Borom induces also enlargement of tubular diameter, epithelial height and tubular lumen. Therefore, it is concluded that Boron acts as testicular toxicant and that further studies are needed to establish its mechanism of action upon spermatogenesis.

El Boro es un elemento químico ampliamente usado en variadas actividades industriales. La exposición a éste afecta a varios órganos en mamíferos, principalmente órganos reproductivos. Este trabajo evaluó los efectos del Boro en testículo. Para este propósito, 12mg Boro/Ide agua potable, equivalente al agua bebestible de Arica, se administró por 42 días a ratones machos CF1 de 85 días de edad (grupo experimental). Otro grupo (grupo control) bebió agua de Santiago (0.6mg Boro/L). Se realizaron estudios histopatológicos y morfométricos del testículo. Estos estudios mostraron que el Boro induce alteraciones como vacuolización epitelial, taponamiento y atrofia del lumen testicular. Los estudios morfométricos mostraron que el Boro también induce aumento del diámetro tubular, altura del epitelio y del lumen tubular. En consecuencia, se concluye que el Boro actúa como un tóxico testicular y que futuros estudios son necesarios para establecer su mecanismo de acción sobre la espermatogénesis.

Testosterone production and spermatogenic damage induced byorganophosphorate pesticides

Parathion is an organophosphorate pesticide amply used in agriculture. Many alterations induced by organophosphorate pesticides have been described, such as: cytogenetic alterations in germinal cells, oligozoospermia and teratozoospermia in the mouse. The effect of Parathion, both pure (PP) and commercial (PC), on mouse interstitial cell testosterone production was evaluated in vivo and in vitro. Male mice were intraperitoneally injected with a single dose of 1/3 LD50 of Parathion, both PP and PC. The animals were sacrificed at 1, 8 and 40 days post injection to evaluate the impact of disrupting testosterone production on spermatogonia, spermatocytes and elongated spermatids. The plasma testosterone was assayed by standard radioimmunoanalysis. The same method was used to assay testosterone in the culture medium of interstitial cells obtained from the control and Parathion treated animals at the same time intervals. Sperm count, sperm teratozoospermia and tubular blockage were analyzed for an appraisal of spermatogenesis. Increase in the teratozoospermia and tubular blockage was detected in the PP and PC group at 8 and 40 days post injection. Plasma testosterone levels drop significantly at 8 days and recovered slowly at 40 days only in PP animals as detected in vivo, implying interference of testicular steroidogenesis due to the toxicant. Recuperation of normality occurs at long time intervals. In conclusion, Parathion disturbs the synthesis of testosterone in mice affecting qualitatively the spermatogenesis.

Testosterone production and spermatogenic damage induced byorganophosphorate pesticides

Parathion is an organophosphorate pesticide amply used in agriculture. Many alterations induced by organophosphorate pesticides have been described, such as: cytogenetic alterations in germinal cells, oligozoospermia and teratozoospermia in the mouse. The effect of Parathion, both pure (PP) and commercial (PC), on mouse interstitial cell testosterone production was evaluated in vivo and in vitro. Male mice were intraperitoneally injected with a single dose of 1/3 LD50 of Parathion, both PP and PC. The animals were sacrificed at 1, 8 and 40 days post injection to evaluate the impact of disrupting testosterone production on spermatogonia, spermatocytes and elongated spermatids. The plasma testosterone was assayed by standard radioimmunoanalysis. The same method was used to assay testosterone in the culture medium of interstitial cells obtained from the control and Parathion treated animals at the same time intervals. Sperm count, sperm teratozoospermia and tubular blockage were analyzed for an appraisal of spermatogenesis. Increase in the teratozoospermia and tubular blockage was detected in the PP and PC group at 8 and 40 days post injection. Plasma testosterone levels drop significantly at 8 days and recovered slowly at 40 days only in PP animals as detected in vivo, implying interference of testicular steroidogenesis due to the toxicant. Recuperation of normality occurs at long time intervals. In conclusion, Parathion disturbs the synthesis of testosterone in mice affecting qualitatively the spermatogenesis.(AU)

Testosterone production and spermatogenic damage induced by organophosphorate pesticides.

Parathion is an organophosphorate pesticide amply used in agriculture. Many alterations induced by organophosphorate pesticides have been described, such as: cytogenetic alterations in germinal cells, oligozoospermia and teratozoospermia in the mouse. The effect of Parathion, both pure (PP) and commercial (PC), on mouse interstitial cell testosterone production was evaluated in vivo and in vitro. Male mice were intraperitoneally injected with a single dose of 1/3 LD50 of Parathion, both PP and PC. The animals were sacrificed at 1, 8 and 40 days post injection to evaluate the impact of disrupting testosterone production on spermatogonia, spermatocytes and elongated spermatids. The plasma testosterone was assayed by standard radioimmunoanalysis. The same method was used to assay testosterone in the culture medium of interstitial cells obtained from the control and Parathion treated animals at the same time intervals. Sperm count, sperm teratozoospermia and tubular blockage were analyzed for an appraisal of spermatogenesis. Increase in the teratozoospermia and tubular blockage was detected in the PP and PC group at 8 and 40 days post injection. Plasma testosterone levels drop significantly at 8 days and recovered slowly at 40 days only in PP animals as detected in vivo, implying interference of testicular steroidogenesis due to the toxicant. Recuperation of normality occurs at long time intervals. In conclusion, Parathion disturbs the synthesis of testosterone in mice affecting qualitatively the spermatogenesis

Melatonin as protective agent for the cytotoxic effects of diazinon in the spermatogenesis in the earthworm Eisenia foetida.

Diazinon (D) is an organophosphorate synthetic insecticide widely used in the world. It inhibits acetylcholinesterase activity and damages reproduction as well as other organic functions mostly by increasing lipid peroxidation. Melatonin (M) is an indolamine secreted by the pineal gland. It performs numerous functions but recently it has been proposed as a good scavenger of oxygen radicals. The earthworm E. foetida is reputed as an excellent bioindicator of environmental chemical pollution. The testicular toxic effect of D and the protective role of M was analyzed in adult E. foetida, at 1, 7, 10, 15 and 30 days after exposure to 1/4, 1/2 and 3/4 of LD50. Sperm counts and the diameter of the seminal receptacles and of their lumina were altered in D exposed worms, which in addition have a lower percentage of survival, decreased weight and show cholinergic effect (coiling of the tail). All these changes were prevented fully or in part by simultaneous exposure to M. The observations confirm that D is a general and testicular toxicant for E. foetida, a good sentinel indicator and stresses the role of M as a protective agent.

Función espermática post-hipoxia simulada en ratón / Simulated post-hypoxia spermatic fuction in mice

Se propone evaluar los cambios reproductivos en ratones sometidos a hipoxia-normoxia simulando condiciones laborales de la faena minera en el norte de Chile. Se estudió hipoxia hipobárica en cámara hipobárica, simulando 4.100 msnm, en tres grupos experimentales y un grupo control (500 msnm, altura promedio de Santiago), estando cada grupo integrado por seis animales. Ratones (Mus musculus) machos adultos fueron sometidos a regímenes de hipoxia (H)-normoxia (N) los siguientes días: 4H; 8H y 4H-4N-4H (4x4x4) y el grupo control en normoxia continua. Se evaluaron: hematocrito, peso testicular y epididimario, y recuento espermático total en testículo y cauda epididimaria. El peso testicular disminuye a los ocho días de exposición hipóxica hipobárica y se recupera con intervalos de cuatro días de normoxia, al igual que el recuento de espermatozoides en cauda; sin embargo, este valor permanece bajo ocho días post-hipoxia. La producción espermática permanece baja en todos los grupos experimentales. Dado que la alternancia 4x4x4 días se utiliza como régimen habitual de trabajo en las faenas mineras chilenas, es imprescindible estudiar los efectos de la altura sobre la fertilidad humana en estas condiciones.

Effects of chronic hypobaric hypoxia on testis histology and round spermatid oxidative metabolism.

In order to evaluate the effects of the exposition to continuous chronic hypobaric hypoxia (CCHH) and intermittent chronic hypobaric hypoxia (ICHH) on testis histology and on oxidative metabolism of spermatogenic cells (SC), male rats were exposed to a 4600-m simulated altitude (PO2: 89.6 mmHg). After 60 days, ICHH and CCHH groups presented a significant decrease in testicular mass, an increase in interstitial space, a decrease in height of the seminiferous epithelium, depletion of cellular elements, vacuolization in epithelial cells and folding of the basal membrane. Round spermatids from animals exposed to CCHH presented a significant decrease in energy-dependent cell shape changes. Round spermatid mitochondria of CCHH rats seem to be limited in their ability to handle reducing equivalents. These mitochondria also appear to be uncoupled under basal conditions. Round spermatids from CCHH rats evidence large oxygen consumption (QO2) insensitive to inhibition by cyanide, a process that could be partly related to lipoperoxidation. Thus, exposure of male rats to CCHH and ICHH induced evident changes in testicular morphology and loss of spermatogenic cells, in all stages of the spermatogenic cycle. This post-meiotic spermatogenic cell loss in the testis correlated well with metabolic changes in round spermatids that evidenced a strong metabolic stress in these cells.

Update on the impact of Chlamydia trachomatis infection on male fertility.

With approximately 90 million cases annually, infection with Chlamydia trachomatis is the most prevalent sexually transmitted bacterial disease in the world. Considering that these infections are often asymptomatic and cause major complications like acute pelvic inflammatory disease, ectopic pregnancy, infertility or infant pneumonia, the estimated costs for diagnosis and treatment in the USA amounts to 2.2 million US dollars for each 500 cases. Therefore, there is a high need for correct, quick and cost-effective diagnosis and treatment of this urogenital tract infection. New innovative therapies provide good results with regard to efficacy and patients' compliance. The success rates of treatments are at least 95%. However, the occurrence of antibiotic resistance should not be ignored and new treatment schemes must be developed. The state-of-the-art of diagnosis and treatment of chlamydial infections as well as the pathophysiology is discussed in this review. In conclusion, infections with C. trachomatis is an important public health problem, especially in third world and developing countries, and more socio-economic studies linking secondary prevention of chlamydial infections, infertility and adverse pregnancy outcome are needed to understand more of its aetiology. In addition, diagnosis and treatment should be improved. Data in men revealed that past infections but not present infections are more related to male infertility. There is still controversial results. In future studies, function of the seminal vesicles and evaluation of the antioxidant capacity should be taken into account when role of C. trachomatis infection on male fertility is assessed.

Serum reproductive hormone levels and sperm production in male adult rats after treatment with arresting, a fraction obtained from seminiferous tubules conditioned medium.

This study used seminiferous tubule (ST) segments from adult rats to condition culture medium that had been concentrated, size fractioned and administered 10-84 days to adult rats by subcutaneous or intratesticular injection and the effects on testes weight, testosterone, luteinizing hormone (LH) and FSH levels and (homogenization-resistant) epididymal sperm count were determined. The conditioned medium obtained 2 days after culture of ST was fractionated in a 30-100 kDa component. The fraction was injected subcutaneously or intratesticularly. This factor(s), named arresting, decreases sperm count in the epididymis from 13 days to 84 days of treatment without changes in serum LH or testosterone levels. The results of the present study suggest that arresting acts on spermiogenesis/spermiation and/or the entry of sperm into the epididymis from the efferent ductules.

Alpaca semen characteristics previous to a mating period.

Increasing the knowledge of the semen characteristics in the alpaca will contribute to understanding one of the many factors that affect the poor fertility rate in this species. Ten adult male alpacas, 2.6-10 years of age, average weight 64.7 +/- 4.7 kg were used. The animals were distributed randomly into two groups of five each and submitted alternatively to two semen collections, using an artificial vagina and sexually receptive females. For the first semen collection the animals had a sexual rest period of about 90 and 45 days before the second. Duration of semen collection, color and volume of ejaculate were recorded, and sperm concentration and morphology (light microscopy) were evaluated. Descriptive statistical analyses were used for each variable, considering all samples obtained (n= 19). An analysis of variance for animal groups and opportunity of collection were used for quantitative variables. Most frequent color was opalescent white (84.2%). There were no statistical differences among male groups or between semen collections. The average values and standard deviations for the quantitative variables were: 12.3 +/- 7.2 min for semen collection time, 1.8 +/- 0.8 ml for ejaculate volume, (17.6 +/- 26.1) x 10(6) sperm/ml for sperm concentration and 34.0 +/- 52.2 x 10(6) for total number of sperm per ejaculate. The percentage of normal spermatozoa was 51.0 +/- 12.4%. From the total abnormalities, that of mid piece segment (14.4%) was the most frequent. These results indicate that male alpaca have poor semen quality, when compared with other domestic species. Nevertheless, for the evaluation of male alpaca as breeders it would be necessary to create a protocol for the selection of them, where phenotypic, behavioral and seminogram aspects are considered. The values reported herein define the characteristics of the alpaca semen that could be considered as the initial base of the seminal analysis to select male alpacas before mating.

Morphometric characterization and classification of alpaca sperm heads using the sperm-class analyzer computer-assisted system.

Sperm morphology has been identified as one characteristic which can be useful in the prediction of sperm fertility, therefore, we hope that this study aimed at establishing standardized morphological criteria might serve in future studies dealing with the search for sperm parameters which facilitate an estimation of sperm quality. For this purpose, ejaculates from fertile alpacas were used to evaluate sperm head morphometry by means of the Sperm-Class Analyzer (SCA) computer-aided image analysis system. We defined three morphological categories according to sperm head size (normal 50%, small 26%, large 24%) and five categories according to sperm head shape (normal 47%, pyriform 3%, short 20%, round 1%, long 29%). Sperm classification according to shape was performed by first morphometrically characterizing sperm heads clearly falling into each of the shape categories. Thereafter, discriminant analysis was performed on the data from these typical sperm heads and the resulting classification functions were used to categorize 2,200 spermatozoa from 11 alpacas. Classification of sperm heads by this method agreed in 88% of the cases with most of the misclassifications being due to pyriform heads classified as long heads. Morphometric values obtained from samples of 50, 100, 150, 175 and 200 sperm heads were compared. At least 150 sperm heads should be evaluated to overcome sample size influence on sperm measurements. Significant differences in sperm morphometry were found between individuals (CV for morphometric parameters ranging from 1.3 to 13.0) and there were marked differences in the sperm morphological composition of the ejaculates. Within-animal CV ranged from 4.7 to 17.8 thus showing the high degree of sperm polymorphism present in the alpaca ejaculate.

Parathion induces mouse germ cells apoptosis.

Germ cell loss occurs in normal spermatogenesis at defined stages of the seminiferous epithelial cycle. The process has been known for over a century but only recently it was analyzed under the concept of apoptosis. This is a programmed cell death that occurs during development and also in the adult. It is believed to play a key role as quality control in sperm formation, avoiding the passage of genetic defects to future generations. Chemical toxicants may increase apoptosis, disturbing tissue homeostasis. The effect of the agropesticide parathion upon apoptosis in mouse seminiferous tubules was analyzed in young mice (onset of spermatogenesis) and in adult animals (full spermatogenesis). In both young and adult mice, the pesticide increases the rate of apoptosis, which takes place at stages where spermatogonial proliferation occurs, affects spermatocytes at the beginning of the meiotic process and spermatids at the elongating period. Basal apoptotic rates are greater in young mice. In adults, commercial parathion is more toxic than the pure organophosphoric compound. From these observations plus in vitro effects of parathion reported previously, it can be concluded that the pesticide affects DNA (and RNA and protein) synthesis. The effect is reversible with moderate doses of the chemical after acute intoxication.

Morphology of the ventral lobe of the prostate and seminal vesicles in an ethanol-drinking strain of rats (UChA and UChB).

Chronic alcoholism alters reproduction and therefore may be responsible for alterations of prostate and seminal vesicles, which are the subject of this analysis in UCh ethanol-drinking rats. The prostate and seminal vesicles of 20 animals were submitted to macroscopic, light microscopy, electron microscopy and morphometric analysis. The UCh rats showed atrophy of the epithelium and reduction of the weight of the prostate and seminal vesicle, liver hypertrophy and fat infiltration and alterations of the hypothalamus-pituitary axis. Ethanol induces changes in the weight and in the epithelium of prostate and seminal vesicles and hypothalamus-pituitary axis of UCh rats.