Abstract: The aim of this study was to evaluate the dose- and time-dependent in vitro effects of berberine (BER), a natural alkaloid with numerous biological properties on bovine spermatozoa during three time periods (0 h, 2 h, 24 h). Bovine semen samples were diluted and cultivated in physiological saline solution containing 0.5% DMSO together with 200, 100, 50, 10, 5, and 1 μmol/L BER. Spermatozoa motility was assessed using the computer assisted semen analyzer. The viability of spermatozoa was assessed by the metabolic (MTT) assay, production of superoxide radicals was quantified using the nitroblue tetrazolium (NBT) test, and chemiluminescence was used to evaluate the generation of reactive oxygen species (ROS). Cell lysates were prepared and the extent of lipid peroxidation (LPO) was evaluated using the TBARS assay. The results of the movement activity showed a significant increase in the motility during long term cultivation in case of concentrations ranging between 1 and 10 μmol/L BER (P < 0.01; P < 0.001; 24 h). At the same time, supplementation of 1, 5 and 10 μmol/L BER led to a significant preservation of the cell viability (P < 0.001; 24 h). BER addition at a range of 1-50 μmol/L also provided a significantly higher protection against superoxide (P < 0.05) and ROS (P < 0.001; P < 0.01) overgeneration as well as LPO (P < 0.01; P
Abstract: In order to improve the animal protein recommended daily intake of Nigerians, there is an upsurge in breeding of hitherto shunned food animals one of which is the rabbit. Radiography and ultrasonography are tools for diagnosing disease and evaluating the anatomical architecture of parts of the body non-invasively. As the rabbit is becoming a more important food animal, to achieve improved breeding of these animals, the best of the species form a breeding stock and will usually depend on breeding soundness which may be evaluated by assessment of the male reproductive organs by these tools. Four male intact rabbits weighing between 1.2 to 1.5 kg were acquired and acclimatized for 2 weeks. Dorsoventral views of the testes were acquired using a digital radiographic machine and a 5 MHz portable ultrasound scanner was used to acquire images of the testes in longitudinal, sagittal and transverse planes. Radiographic images acquired revealed soft tissue images of the testes in all rabbits. The testes lie in individual scrotal sacs sides on both sides of the midline at the level of the caudal vertebrae and thus are superimposed by caudal vertebrae and the caudal limits of the pelvic girdle. The ultrasonographic images revealed mostly homogenously hypoechogenic testes and a hyperechogenic mediastinum testis. The dorsal and ventral poles of the testes were heterogeneously hypoechogenic and correspond to the epididymis and spermatic cord. The rabbit is unique in the ability to retract the testes particularly when stressed and so careful and stressless handling during the procedures is of paramount importance. The imaging of rabbit testes can be safely done using both imaging methods but ultrasonography is a better method of assessment and evaluation of soundness for breeding.
Abstract: Main sources of endocrine disrupting compounds in the ecosystem are hormones, pesticides, phthalates, flame retardants, dioxins, personal-care products, coplanar polychlorinated biphenyls (PCBs), bisphenol A, and parabens. These endocrine disrupting compounds are responsible for learning disabilities, brain development problems, deformations of the body, cancer, reproductive abnormalities in females and decreased sperm count in human males. Although discharge of these chemical compounds into the environment cannot be stopped, yet their amount can be retarded through proper evaluation and detection techniques. The available techniques for determination of these endocrine disrupting compounds mainly include high performance liquid chromatography (HPLC), mass spectroscopy (MS) and gas chromatography-mass spectrometry (GC–MS). These techniques are accurate and reliable but have certain limitations like need of skilled personnel, time consuming, interference and requirement of pretreatment steps. Moreover, these techniques are laboratory bound and sample is required in large amount for analysis. In view of above facts, new methods for detection of endocrine disrupting compounds should be devised that promise high specificity, ultra sensitivity, cost effective, efficient and easy-to-operate procedure. Nowadays, electrochemical sensors/biosensors modified with nanomaterials are gaining high attention among researchers. Bioelement present in this system makes the developed sensors selective towards analyte of interest. Nanomaterials provide large surface area, high electron communication feature, enhanced catalytic activity and possibilities of chemical modifications. In most of the cases, nanomaterials also serve as an electron mediator or electrocatalyst for some analytes.
Abstract: The present study was carried out on Sahiwal cattle bulls maintained at the Artificial Breeding Complex, NDRI, Karnal, Hayana, India, to assess the effect of cooling using mist cooling and fanning on Sahiwal bulls in the dry hot summer season. Fourteen Sahiwal bulls were divided into two groups of seven each. Sexual behavior and semen quality traits considered were: Reaction time (RT), Dismounting time (DMT), Total time taken in mounts (TTTM), Flehmen response (FR), Erection Score (ES), Protrusion Score (PS), Intensity of thrust (ITS), Temperament Score (TS), Libido Score (LS), Semen volume, Physical appearance, Mass activity, Initial progressive motility, Non-eosinophilic spermatozoa count (NESC) and post thaw motility percent. Data were analyzed by least squares technique. Group-1 was the control, whereas group-2 (treatment group) bulls were exposed to mist cooling and fanning (thrice a day 15 min each) in the dry hot summer season. Group-2 showed significantly (p < 0.01) higher value in DMT (sec), ES, PS, ITS, LS, semen volume (ml), semen color density, mass activity, initial motility, progressive motility and live sperm.
Abstract: Mechanical index (MI) is used for quantifying acoustic cavitation and the relationship between acoustic pressure and the frequency. In this study, modeling of the MI was applied to provide treatment protocol and to understand the effective physical processes on reproducibility of stem cells. The acoustic pressure and MI equations are modeled and solved to estimate optimal MI for 28, 40, 150 kHz and 1 MHz frequencies. Radial and axial acoustic pressure distribution was extracted. To validate the results of the modeling, the acoustic pressure in the water and near field depth was measured by a piston hydrophone. Results of modeling and experiments show that the model is consistent well to experimental results with 0.91 and 0.90 correlation of coefficient (p
Abstract: The study was conducted on Sahiwal cattle bulls maintained at the Artificial Breeding Complex, NDRI, Karnal, Hayana, India, to determine the effect of exercise on the sexual behavior and semen quality. Fourteen Sahiwal bulls were classified into two groups of seven each. Group-1, bulls were exercised by walking in a bull exerciser once a week one hour before semen collection, whereas bulls in group-2 were exercised daily. Sexual behavior and semen quality traits studied were: Reaction time (RT), Dismounting time (DMT), Total time taken in mounts (TTTM), Flehmen response (FR), Erection Score (ES), Protrusion Score (PS), Intensity of thrust (ITS), Temperament Score (TS), Libido Score (LS), Semen volume, Physical appearance, Mass activity, Initial progressive motility, Non-eosinophilic spermatozoa count (NESC) and post thaw motility percent. Data were analyzed by least squares technique. Group-2 showed significantly (p < 0.01) higher value in RT (sec), DMT (sec), TTTM (sec), ES, PS, ITS, LS, semen volume, semen color density and mass activity.
Abstract: This study aimed to assess the in vitro effects of different concentrations of the Viscum album extract on the motility, viability, and reactive oxygen species (ROS) production by rabbit spermatozoa during different time periods (0, 2, and 8h). Spermatozoa motility was assessed by using the CASA (Computer aided sperm analysis) system. Cell viability was evaluated by using the metabolic activity MTT assay, and the luminol-based luminometry was applied to quantify the ROS formation. The CASA analysis revealed that low Viscum concentrations were able to prevent a rapid decline of spermatozoa motility, especially in the case of concentrations ranging between 1 and 5 µg/mL (P
Abstract: In vitro storage and processing of animal semen represents a risk factor to spermatozoa vitality, potentially leading to reduced fertility. A variety of substances isolated from natural sources may exhibit protective or antioxidant properties on the spermatozoon, thus extending the lifespan of stored ejaculates. This study compared the ability of different concentrations of the Salvia officinalis extract on the motility, mitochondrial activity, viability and reactive oxygen species (ROS) production by bovine spermatozoa during different time periods (0, 2, 6 and 24 h) of in vitro culture. Spermatozoa motility was assessed using the Computer-assisted sperm analysis (CASA) system. Cell viability was examined using the metabolic activity MTT assay, the eosin-nigrosin staining technique was used to evaluate the sperm viability and ROS generation was quantified using luminometry. The CASA analysis revealed that the motility in the experimental groups supplemented with 0.5-2 µg/mL Salvia extract was significantly lower in comparison with the control (P
Abstract: Snails are considered as suitable diagnostic organisms for heavy metal–contaminated sites. Biomphalaria alexandrina snails are used in this work as pollution bioindicators after exposure to chemical mixtures consisted of heavy metals (HM); zinc (Zn), copper (Cu) and lead (Pb); and persistent organic pollutants; Decabromodiphenyl ether 98% (D) and Aroclor 1254 (A). The impacts of these tested chemicals, individual and mixtures, on liver and kidney functions, antioxidant enzymes, complete blood picture, and tissue histology were studied. Results showed that Cu was proved to be the highly toxic against snails than Zn and Pb where LC50 values were 1.362, 213.198 and 277.396 ppm, respectively. Also, B. alexandrina snails exposed to the mixture of HM (¼ LC5 Cu, Pb and Zn) showed the highest bioaccumulation of Cu and Zn in their whole tissue, the most significant increase in AST, ALT & ALP activities and the highest significant levels of total protein, albumin and globulin. Results showed significant alterations in CAT activity in snail tissue extracts while snail samples exposed to most experimental tests showed significant increase in GST activity. Snail samples that exposed to HM mixtures showed a significant decrease in total hemocytes count while snail samples that exposed to mixtures containing A & D showed a significant increase in total hemocytes and Hyalinocytes. Histopathological alterations in snail samples exposed to individual HM and their mixtures for 4 weeks showed degeneration, edema, hyper trophy and vaculation in head-foot muscle, degeneration and necrotic changes in the digestive gland and accumulation in most tested organs. Also, the hermaphrodite gland showed mature ova with irregular shape and reduction in sperm number. In conclusion, the resulted damage and alterations in B. alexandrina studied parameters can be used as bioindicators to the presence of pollutants in its habitats.
Abstract: The present in vitro study was designed to reveal whether amygdalin (AMG) is able to cause changes to the motility, viability and mitochondrial activity of rabbit spermatozoa. New Zealand White rabbits (n = 10) aged four months were used in the study. Semen samples were collected from each animal and used for the in vitro incubation. The samples were divided into five equal parts and diluted with saline supplemented with 0, 0.5, 1, 2.5 and 5 mg/mL AMG. At times 0h, 3h and 5h spermatozoa motion parameters were assessed using the SpermVision™ computer-aided sperm analysis (CASA) system, cell viability was examined with the metabolic activity (MTT) assay, and the eosin-nigrosin staining technique was used to evaluate the viability of rabbit spermatozoa. All AMG concentrations exhibited stimulating effects on the spermatozoa activity, as shown by a significant preservation of the motility (P
Abstract: Oxidative stress associated with semen cryopreservation may result in lipid peroxidation (LPO), DNA damage and apoptosis, leading to decreased sperm motility and fertilization ability. Curcumin (CUR), a natural phenol isolated from Curcuma longa Linn. has been presented as a possible supplement for a more effective semen cryopreservation because of its antioxidant properties. This study focused to evaluate the effects of CUR on selected oxidative stress parameters in cryopreserved bovine semen. 20 bovine ejaculates were split into two aliquots and diluted with a commercial semen extender containing CUR (50 μmol/L) or no supplement (control), cooled to 4 °C, frozen and kept in liquid nitrogen. Frozen straws were thawed in a water bath for subsequent experiments. Computer assisted semen analysis was used to evaluate spermatozoa motility, and reactive oxygen species (ROS) generation was quantified by using luminometry. Superoxide generation was evaluated with the NBT test, and LPO was assessed via the TBARS assay. CUR supplementation significantly (P
Abstract: This work investigated possible inductions of CaC2, often misused by fruit vendors to stimulate artificial ripening, on mammalian sperm morphology and viability. Thirty isogenic strains of male albino mice, Mus musculus (age≈ 8weeks; weight= 32.52.0g) were acclimatized (ambient temperature 28.0±1.0°C) for 2 weeks and fed standard growers mash and water ad libutum. They were later exposed to graded toxicant concentrations (w/w) of 2.5000, 1.2500, 0.6250, and 0.3125% in 4 cages. A control cage was also established. After 5 weeks, 3 animals from each cage were sacrificed by cervical dislocation and the cauda epididymis excised. Sperm morphology and viability were determined by microscopic procedures. The ANOVA, means plots, Student’s t-test and variation plots were used to analyze data. The common abnormalities observed included Double Head, Pin Head, Knobbed Head, No Tail and With Hook. The higher toxicant concentrations induced significantly lower body weights [F(829.899) ˃ Fcrit(4.19)] and more abnormalities [F(26.52) ˃ Fcrit(4.00)] at P˂0.05. Sperm cells in the control setup were significantly more viable than those in the 0.625% (t=0.005) and 2.500% toxicant doses (t=0.018) at the 95% confidence limit. CaC2 appeared to induced morphological abnormalities and reduced viability in sperm cells of M. musculus.
Abstract: This research work is an experimental study, through
development of an adhesive from Prosopis africana endosperm. The
prosopis seed for this work were obtained from Enugu State in the
South East part of Nigeria. The seeds were prepared by separating the
endosperm from the seed coat and cotyledon. Three methods were
used to separate them, which are acidic method, roasting method and
boiling method. 20g of seed were treated with different
concentrations (25, 40, 55, 70, and 85% w/w) at 100°C and constant
time (30 minutes), under continuous stirring with magnetic stirrer.
Also 20g of seed were treated with sulphuric acid of concentrations
40% w/w at 100°C with different time (10, 15, 20, 25, 30 minutes),
under continuous stirring with magnetic stirrer. Finally, 20g of seed
were treated with sulphuric acid of concentrations 40% w/w at
different temperature (20°C, 40°C, 60°C, 80°C, and 100°C) with
constant time (30 minutes), under continuous stirring with magnetic
stirrer. The whole endosperm extracted was adhesive. The physical
properties of the adhesive were determined (appearance, odour, taste,
solubility, pH, size, and binding strength). The percentage of the
adhesive yield makes the commercialization of the seed in Nigeria
possible and profitable. The very high viscosity attained at low
concentrations makes prosopis adhesive an excellent thickener in the
food industry.
Abstract: Wheat is the first and the most important grain of the
world and its bakery property is due to glutenin and gliadin qualities.
Wheat seed proteins were divided into four groups according to
solubility including albumin, globulin, glutenin and prolamin or
gliadin. Gliadins are major components of the storage proteins in
wheat endosperm. It seems that little information is available about
gliadin genes in Iranian wild relatives of wheat. Thus, the aim of this
study was the evaluation of the wheat wild relatives collected from
different origins of Zagros Mountains in Iran, in terms of coding
gliadin genes using specific primers. For this, forty accessions of
Triticum boeoticum and Triticum urartu were selected for this study.
For each accession, genomic DNA was extracted and PCRs were
performed in total volumes of 15 μl. The amplification products were
separated on 1.5% agarose gels. In results, for Gli-2A locus three
allelic variants were detected by Gli-2As primer pairs. The sizes of
PCR products for these alleles were 210, 490 and 700 bp. Only five
(13%) and two accessions (5%) produced 700 and 490 bp fragments
when their DNA was amplified with the Gli.As.2 primer pairs.
However, 93% of the accessions carried allele 210 bp, and only 8%
did not any product for this marker. Therefore, these germplasm
could be used as rich gene pool to broaden the genetic base of bread
wheat.
Abstract: Target of this study was the analysis of the impact of
crude glycerol on canine spermatozoa motility, morphology,
viability, and membrane integrity. Experiments were realized in vitro.
In the study, semen from 5 large dog breeds was used. They were
typical representatives of large breeds, coming from healthy rearing,
regularly vaccinated and integrated to the further breeding. Semen
collections were realized at the owners of animals and in the
veterinary clinic. Subsequently the experiments were realized at the
Department of Animal Physiology of the SUA in Nitra. The
spermatozoa motility was evaluated using CASA analyzer
(SpermVisionTM, Minitub, Germany) at the temperature 5 and 37°C
for 5 hours. In the study, 13 motility parameters were evaluated.
Generally, crude glycerol has generally negative effect on
spermatozoa motility. Morphological analysis was realized using
Hancock staining and the preparations were evaluated at
magnification 1000x using classification tables of morphologically
changed spermatozoa. Data clearly detected the highest number of
morphologically changed spermatozoa in the experimental groups
(know twisted tails, tail torso and tail coiling). For acrosome
alterations swelled acrosomes, removed acrosomes and acrosomes
with undulated membrane were detected. In this study also the effect
of crude glycerol on spermatozoa membrane integrity were analyzed.
The highest crude glycerol concentration significantly affects
spermatozoa integrity. Results of this study show that crude glycerol
has effect of spermatozoa motility, viability, and membrane integrity.
Detected changes are related to crude glycerol concentration,
temperature, as well as time of incubation.
Abstract: More than 3000 plants of notable phyto-therapeutic
value grow in South Africa; these include Cissampelos capensis,
commonly known in Afrikaans as dawidjie or dawidjiewortel. C.
capensis is the most significant and popular medicinal plant used by
the Khoisan as well as other rural groups in the Western region of
South Africa. Its rhizomes are traditionally used to treat male fertility
problems. Yet, no studies have investigated the effects of this plant or
its extracts on human spermatozoa. Therefore, this study aimed at
investigating the effects of C. capensis rhizome extract (CRE)
fractions on ejaculated human spermatozoa in vitro. Spermatozoa
from a total of 77 semen samples were washed with human tubular
fluid medium supplemented with bovine serum albumin (HTF-BSA)
and incubated for 2 hours with 20 μg/ml progesterone (P4) followed
by incubation with different concentrations (0, 0.05, 0.5, 5, 50, 200
μg/ml) of fractionated CRE (F1=0% MeOH, F2=30% MeOH,
F3=60% MeOH and F4=100% MeOH) for 1.5 hours at 37°C. A
sample without addition of CRE fractions served as control. Samples
were analyzed for sperm motility, reactive oxygen species (ROS),
DNA-fragmentation, acrosome reaction and capacitation. Results
showed that F1 resulted in significantly higher values for ROS,
capacitation and hyper-activation compared to F2, F3, and F4 with
P4-stimulated samples generally having higher values. No significant
effect was found for the other parameters. In conclusion, alkaloids
present in F1 of CRE appear to have triggered sperm intrinsic ROS
production leading to sperm capacitation and acrosome reaction
induced by P4.
Abstract: The influences of cell-free solutions (CFSs) of lactic
acid bacteria (LAB) on cadaverine and other biogenic amines
production by Listeria monocytogenes and Staphylococcus aureus
were investigated in lysine decarboxylase broth (LDB) using HPLC.
Cell free solutions were prepared from Lactococcus lactis subsp.
lactis, Leuconostoc mesenteroides subsp. cremoris, Pediococcus
acidilactici and Streptococcus thermophiles. Two different
concentrations that were 50% and 25% CFS and the control without
CFSs were prepared. Significant variations on biogenic amine
production were observed in the presence of L. monocytogenes and S.
aureus (P < 0.05). The function of CFS on biogenic amine production
by foodborne pathogens varied depending on strains and specific
amine. Cadaverine formation by L. monocytogenes and S. aureus in
control were 500.9 and 948.1 mg/L, respectively while the CFSs of
LAB induced 4-fold lower cadaverine production by L.
monocytogenes and 7-fold lower cadaverine production by S. aureus.
The CFSs resulted in strong decreases in cadaverine and putrescine
production by L. monocytogenes and S. aureus, although remarkable
increases were observed for histamine, spermidine, spermine,
serotonin, dopamine, tyramine and agmatine in the presence of LAB
in lysine decarboxylase broth.
Abstract: The presence of endocrine-disrupting compounds,
such as bisphenol A (BPA), in the environment can cause serious
health problems. However, there are controversial opinions. This
study investigated the reproductive, metabolic, oxidative and
immunologic-disrupting effects of bisphenol A in male rabbits.
Rabbits were divided into five groups. The first four rabbit groups
were administered oral BPA (1, 10, 50, or 100 mg/kg/day) for ten
weeks. The fifth group was administered corn oil as the vehicle. BPA
significantly decreased serum testosterone, estradiol and the free
androgen index (FAI) and significantly increased sex hormone
binding globulin (SHBG) compared with the placebo group. The
higher doses of BPA showed a significant decrease in follicular
stimulating hormone (FSH) and luteinizing hormone (LH). A
significant increase in blood glucose levels was identified in the BPA
groups. The non-significant difference in insulin levels is a novel
finding. The cumulative testicular toxicity of BPA was clearly
demonstrated by the dose-dependent decrease in absolute testes
weight, primary measures of semen quality and a significant increase
in testicular malonaldehyde (MDA). Moreover, BPA significantly
decreased total antioxidant capacity (TAC) and significantly
increased immunoglobulin G (IgG) at the highest concentration. Our
results suggest that BPA, especially at higher doses, is associated
with many adverse effects on metabolism, oxidative stress, immunity,
sperm quality and markers of androgenic action.
Abstract: Malathion (ML) is a well known pesticide commonly
used in many agricultural and non-agricultural processes. Its toxicity
has been attributed primarily to the accumulation of acetylcholine
(Ach) at nerve junctions, due to the inhibition of acetylcholinesterase
(AChE). The aim of the current research was to study the protective
effect of the melissa plant extract against reproductive impairment
induced by malathion in 32 male albino rats, and the biological
experiment was divided into four groups (8 in each) that given
malathion (27 mg/kg; 1/50 of the LD50 for an oral dose) and/or
Melissa officinalis (MO) extract (200mg/kg/day) by gavages
technique. The sperm counts, sperm motility, sperm morphology,
FSH, LH, and testosterone levels had been determined in testes
homogenate at the end of the experiment. It is worthy to report that,
rats treated with melissa extract did not show a significant difference
when compared with the control group, while rats given malathion
alone had significantly lower sperm count, sperm motility, and
significantly higher abnormal sperm numbers, than the untreated
control rats as well as having significantly lower serum FSH, LH, and
testosterone levels compared with the control group. Administrations
of melissa extract restore all mentioned histological parameters
towards the control group and the melissa extract had a strong
positive protective effect against malathion toxicity. Results the of
biological parameters were confirmed by the histological
examination of rat testes and indicated that, both control and melissa
groups showing normal seminiferous tubules, while malathion group
testicular tissues had necrosis, edema in the seminiferous tubules and
degeneration of spermatogonial cells lining the seminiferous tubules
with incomplete spermatogenesis. The use of melissa against
malathion improved the histological picture and showing normal
seminiferous tubules with complete spermatogenesis and almost there
was no histopathological changes could be noted.
Abstract: Surgical trauma seems to facilitate metastatic spread, although the underlying mechanisms are not known. Increased concentrations of polyamines (spermine and spermidine) in the blood seem to have associated with the enhanced malignant potential of cancer cells and decrease in anti-tumor immunity of cancer patients. In addition to de novo synthesis in rapidly growing cells such as normal regenerating cells and cancer cells, cells can take up polyamines from extra-cellular sources. We have shown that increased polyamine concentration results in decreases in cytokine production and expression of adhesion molecules involved in anti-tumor immunity, such as CD11a. And, immune cells in an environment with increased polyamine levels lose anti-tumor immune functions, such as lymphokine activated killer cell (LAK) activities. Because blood polyamine levels are increased in post-surgical patients, polyamine seems to have roles on post-traumatic tumor spread.