Sucralose Influences the Productive Performance, Carcass Traits, Blood Components, and Gut Microflora Using 16S rRNA Sequencing of Growing APRI-Line Rabbits.

This study investigated how sucralose influenced rabbit intestine and caecal microbial activity, blood parameters, growth performance, carcass characteristics, and digestibility. In total, 160 5-week-old rabbits from the APRI line weighing 563.29 gm were randomly assigned to four experimental groups with four replicates-5 males and 5 females in each. Four experimental groups were used, as follows: SUC1, SUC2, and SUC3 got 75, 150, and 300 mg of sucralose/kg body weight in water daily, while the control group ate a basal diet without supplements. The results showed that both the control and SUC1 groups significantly (p < 0.05) increased daily weight gain and final body weight. Sucralose addition significantly improved feed conversion ratio (p < 0.05) and decreased daily feed intake (gm/d). The experimental groups do not significantly differ in terms of mortality. Furthermore, nutrient digestibility was not significantly affected by sucralose treatment, with the exception of crud protein digestion, which was significantly reduced (p < 0.05). Additionally, without altering liver or kidney function, sucralose administration dramatically (p < 0.05) decreased blood serum glucose and triglyceride levels while increasing total lipids, cholesterol, and malonaldehyde in comparison to the control group. Furthermore, the addition of sucrose resulted in a significant (p < 0.05) increase in the count of total bacteria, lactobacillus, and Clostridium spp., and a decrease in the count of Escherichia coli. Further analysis using 16S rRNA data revealed that sucralose upregulated the expression of lactobacillus genes but not that of Clostridium or E. Coli bacteria (p < 0.05). Therefore, it could be concluded that sucralose supplementation for rabbits modifies gut microbiota and boosts beneficial bacteria and feed conversion ratios without side effects. Moreover, sucralose could decrease blood glucose and intensify hypercholesterolemia and should be used with caution for human consumption.

Impact of dietary Moringa oleifera leaf supplementation on gut morphometry, behaviour and physiological parameters in growing male rabbits.

Moringa oleifera (MO), a cultivated species of the Moringa, is known for its high concentration of essential nutrients that promote growth. To assess its impact on rabbits' gut morphometric, behavioural, and physiological parameters, a study was conducted using sixty growing male white New Zealand rabbits at 40 days old. The rabbits were divided into four groups and supplemented with dried MO leaves at varying levels (0%, 0.5%, 1% and 2% of body weight) for four weeks. The results revealed significant increases in organ weights, such as liver and intestinal length, and the height of intestinal villi and crypt depth in the large intestine. The muscular layer's and submucosa's thickness also increased in different parts of the intestine in rabbits fed with MO compared to the control group. No significant effect was observed on the caecum mucosa depth. Interestingly, no significant differences were found in body weight or weight gain between the Moringa supplementation groups and the control group. However, rabbits offered 0%, 0.5% and 1% MO spent more time feeding and resting than those given 2% MO. The grooming and sniffing percentage remained unaffected by Moringa supplementation. Regarding blood parameters, rabbits that received MO leaves in their diet showed improvements in red blood cell count, haemoglobin levels, packed cell volume, white blood cell count, neutrophil count, total protein, globulin, and A/G ratio. Moreover, there was a significant decrease in malondialdehyde levels and an increase in glutathione and high-density lipoprotein levels, indicating an antioxidative effect. Overall, the study concluded that MO leaves supplementation in the rabbit diet positively influenced the rabbits' health by modulating the immune system, improving histological aspects of the intestine, liver, and spleen, and enhancing physiological parameters through its antioxidative properties.

Evaluation of the Maturity and Gene Expression of Sheep Oocytes and Embryos Cultured in Media Supplemented with Marjoram (Origanum vulgare) Extract.

This study aimed to evaluate the effects of marjoram extract on oocyte maturation and gene expression in sheep oocytes and embryos. The first experiment studied the effect of the extract as an antioxidant to improve the in vitro maturation media used for sheep oocytes; the oocytes were matured in a TCM199 medium supplemented with 1 or 10 µg/mL of marjoram extract or the control, 0 µg, for 24 hr. Then, the maturation was estimated, and the gene expression was measured by using qPCR. The second experiment studied the effect of the extract on the development of sheep embryos produced in vitro; the fertilized oocytes were cultured in a SOF medium supplemented with 1 or 10 µg/mL of marjoram extract or the control, 0 µg, for 7 days. Then, the gene expression was measured using qPCR. The results showed that the marjoram extract did not improve nuclear maturation or the blastocyst rate. There was a significant increase in the level of GDF-9 gene expression in mature oocytes in the treatment groups. An increase in the expression of BCL-2 and EGR-1 genes was observed for the blastocysts in the 10 µg/mL group. We concluded that the marjoram extract did not improve nuclear maturation, but it did affect the expression of some genes in sheep oocytes and embryos.

Ameliorative effect of L-carnitine on lambda-cyhalothrin-induced anatomical and reproductive aberrations in albino mice.

The objective of this research was to look at how the pesticide lambda-cyhalothrin (LCT) affected the liver, kidney, testis, and ovary of albino mice; and on morphological and skeletal features of the newborn of treated females. The study also aimed to test the ameliorative effects of L-carnitine (LC) against (LCT). Five sets of mice were created, Group 1 acted as the control, while Group 2 received a high dose of LCT, Group 3 received a high dose of LCT + LC, Group 4 received a low dose of LCT that was a residue of in khat (Qat), and Group 5 received a low dose of LCT + LC. The findings revealed that the treated groups' body weights were reduced significantly, whereas the absolute and relative weights of the liver in all groups were statistically decreased insignificantly. There were histopathological changes in the tissues in groups 2 and 4. While the tissues of the ovary and testis showed recovery in groups 3 and 5. When compared to the control group, the values of the seminiferous tubules parameters were statistically significant in the 3 and 5 groups. The newborn had a high dose of pesticides and showed some malformations in the skeleton. However, in group 3 the skeletal malformation was minimized and in-group 5 the skeleton malformations had completely disappeared. It could be concluded that LCT is highly harmful to mouse tissues and caused neonatal malformations, whereas LC has a marked protective effect against LCT.

Prevalence of Anaplasma marginale in cattle blood samples collected from two important livestock regions in Punjab (Pakistan) with a note on epidemiology and phylogeny of parasite.

Anaplasmosis, caused by intracellular gram-negative bacteria Anaplasma marginale is one of the most frequently reported tick-borne disease (TBDs) in tropical and sub-tropical countries, including Pakistan. In the present study, a total of 428 cattle blood samples were collected to examine the prevalence and phylogenetic origin of A. marginale in two important livestock regions of Punjab Province in Pakistan, i.e. Lodhran and Dera Ghazi Khan Districts. In addition, association between occurrence of A. marginale in cattle blood and selected epidemiological factors has been also investigated. The presence of A. marginale genetic material was confirmed in 9% of the tested blood samples taken from cattle in Lodhran and in 17% from Dera Ghazi Khan. Prevalence of A. marginale was significantly higher in cattle from Dera Ghazi Khan. All the cattle breeds from both districts were equally susceptible to A. marginale infection. We reported higher prevalence of A. marginale in cattle living indoors or with other dairy animals in Dera Ghazi Khan district. However, no such relationship was observed in the Lodhran district. Sequencing of the msp1b gene shows 96-99% similarity of A. marginale in the study area to those reported from other parts of Pakistan, South Africa, and Israel. We recommend that large scale tick and tick-borne disease control strategies must be implemented in both districts.

Morphological and molecular characterization of Ascaridia columbae in the domestic pigeon (Columba livia domestica) and the assessment of its immunological responses.

Parasitism is a divesting problem that is frequently overlooked and may result in severe prominent clinical manifestation. This study aimed to investigate the seasonal and sexual prevalence of the gastrointestinal nematode Ascaridia columbae (A. columbae) infection among domestic pigeons in Giza governorate, Egypt, during the period from 2020 to 2021. One hundred and sixty suspected pigeons were clinically investigated. Blood & tissue samples were collected from infected birds to estimate serum zinc concentration, malondialdehyde (MDA), and nitric oxide levels. As well as tumor necrosis factor-alpha (TNF-α), interleukin 1ß (IL1ß) activity, and histopathological examination were estimated; also, worms were collected for morphological identification using electron microscope (SEM) and molecularly identified using polymerase chain reaction (PCR), further sequenced, and submitted in GenBank with accession number MZ343369. The average ascarid (length × breadth) were 72.4 ± 3.3 µm (70.5 - 79.9 µm) × 39.9 ± 2.5 µm (37.6 - 42.3 µm). The distinguishing morphological characteristics that have been noticed in ascarid worms were creamy white, cylindrical worm with triradiate lips with wide cephalic alae extending on both the lateral sides and filariform esophagus. In males, spicules were almost equal with the presence of precloacal chitinous-rimmed sucker. The prevalence of A. columbae infection was (63.1%) with a higher incidence in females (79.2%) than males (46.1%). The highest seasonal prevalence was observed in winter (92.5%), followed by summer and spring (87.5% and 55%), respectively while, the lowest prevalence was observed in autumn (17.5%). The intensity of worms in the infected intestine varied from 5 to 120 adult worms. The histopathological examination revealed the presence of chronic diffuse moderate catarrhal enteritis with roundworms in the lumen. Infected birds showed a significant increase in nitric oxide and MDA levels while serum zinc levels were lowered in infected pigeons. Infected pigeons revealed a marked increase in IL1-ß and TNFα than apparently healthy ones.

Interspecific Nuclear Transfer Blastocysts Reconstructed from Arabian Oryx Somatic Cells and Domestic Cow Ooplasm.

Cloning, commonly referred to as somatic cell nuclear transfer (SCNT), is the technique of enucleating an oocyte and injecting a somatic cell into it. This study was carried out with interspecific SCNT technology to clone the Arabian Oryx utilizing the oryx's fibroblast cells and transfer it to the enucleated oocytes of a domestic cow. The recipient oocytes were extracted from the cows that had been butchered. Oryx somatic nuclei were introduced into cow oocytes to produce embryonic cells. The study was conducted on three groups, Oryx interspecific somatic cell nuclear transfer into enucleated oocytes of domestic cows, cow SCNT "the same bovine family species", used as a control group, and in vitro fertilized (IVF) cows to verify all media used in this work. The rates of different embryo developmental stages varied slightly (from 1- cell to morula stage). Additionally, the oryx interspecies Somatic cell nuclear transfer blastocyst developmental rate (9.23%) was comparable to that of cow SCNT (8.33%). While the blastula stage rate of the (IVF) cow embryos exhibited a higher cleavage rate (42%) in the embryo development stage. The results of this study enhanced domestic cow oocytes' ability to support interspecific SCNT cloned oryx, and generate a viable embryo that can advance to the blastula stage.

Risk factor analysis for the prevalence of gastrointestinal parasites found in large ruminants in Lower Dir Khyber Pakhtunkhwa Pakistan.

The present study was designed to investigate the prevalence of gastrointestinal (GI) parasites in cattle and buffaloes of Lower Dir Khyber Pakhtunkhwa, Pakistan. The presence of the eggs, cysts, and oocysts of GI parasites in fecal samples were detected using direct smear methods and concentration techniques including floatation, centrifugation, and sedimentation. Identification of recovered fecal stages were determined by morphology using size and appearance of the recovered eggs, cysts, and oocysts. A total of 314 fecal samples were collected from the different Tehsils (Administrative Districts) and analyzed through microscopy. A higher prevalence was observed in the buffalo than the cow population. A total of 184 samples were positive for GI parasites of which 109/196 (55.61%) were from cattle, whereas 75/118 (63.55%) were from buffaloes. The minimum number of strongyle eggs detected in all the samples were 136.39 eggs/g (EPG). The mean EPG in cattle was 143.30 and 122.56 in buffaloes. The open-source water prevalence of GI parasites was higher than the other sources in cattle and the second highest after tap water in buffaloes. The seasonal prevalence of GI parasites ranged from 32.39% (23/71), in spring to 68.8% (86/125) in summer in cattle. In was For buffaloes the infection prevalence was 52.94% (27/51) and 71.64% (48/67) in spring and summer, respectively. Gastrointestinal parasites are a serious problem in cattle and buffaloes in the lower district of Dir Khyber Pakhtunkhwa Pakistan. In general, the burden of parasitic infection was low in most animals that received previous anti-parasitic treatment.

Impacts of tea tree or lemongrass essential oils supplementation on growth, immunity, carcass traits, and blood biochemical parameters of broilers reared under different stocking densities.

The effects of tea tree essential oil (TTEO) and lemongrass essential oil (LGEO) with different stocking densities on the growth performance, biochemical markers, antioxidants, and immunity state of broiler chickens were studied.  Birds were housed at stocking densities of 25, 30, 35, and 40 kg/m2. The treatments were, basal diet without any supplementation, the second and third groups were supplemented with 300 mg TTEO/kg feed, and 300 mg LGEO/kg feed, respectively. Results revealed that increasing stocking density from 25 to 40 kg/m2 significantly reduced body weight and daily weight gain at different ages. The phagocytic index and activity were significantly higher under the lower stocking density (25 kg/m2). Serum amyloid A (SAA), serum or liver transferrin (TRF), or C-reactive protein (CRP) were significant decreased when decreasing stocking density. Increasing stocking density from 25 to 40 kg/m2 resulted in a significant increase in the serum urea, creatinine, uric acid, lactate dehydrogenase (LDH), alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA), and catalase (CAT) levels. However, there was a significant reduction in antioxidant enzyme activity, including glutathione peroxidase (GPx) and superoxide dismutase (SOD), as stocking density increased. The supplementation of TTEO produced significantly higher body weight and daily weight gain followed by LGEO. Additionally, the mortality rates were reduced in TTEO (27.4%) and LGEO (25%) groups. TTEO or LGEO supplementation significantly improved meat constituents and cellular immunity and reduced serum total lipids, serum and meat cholesterol, and triglycerides, SAA, TRF, and CRP. For all these measured parameters, superior results were obtained when TTEO was used compared to LGEO. TTEO or LGEO supplementation also significantly reduced serum urea, creatinine, uric acid, and the enzymatic activities of LDH, ALT, AST, MDA, and CAT (but not GPx and SOD) in comparison to the control treatment. Overall, our results showed the superiority of TTEO over LGEO as a feed supplement in broiler diets. In conclusion, TTEO treatment offers a better solution for raising broiler chickens in high stocking density.

Potential impacts of COVID-19 on reproductive health: Scientific findings and social dimension.

COVID-19 virus is classified as a respiratory disease that can be mainly transmitted via respiratory droplets, however, there are recently published reports suggested its ability to transmit via sexual intercourse, assisted reproductive technology (ART) treatments, pregnancy, and nursing. Although SARS-CoV-2 mainly attacks respiratory systems, manifestations of multiple organs have been detected. A significant concern was raised about whether COVID-19 may affect female and male reproductive functions. These findings imposed more restrictions on social relationships between individuals even if inside the family, adding more physiologic load. In this context, there is a crucial need to identify the biological and behavioral reproductive risk factors associated with COVID-19 disease. Questions regarding the potential risks of sexual transmission during intercourse and/or application of ART, vertical transmission (throughout pregnancy, delivery, and breastfeeding), the health of pregnant and postpartum women, and fetal or postnatal health problems of neonates/children remain largely unanswered. The contribution of individuals to different social and economic activities depends on the maintenance of good quality life and health. The ongoing COVID-19 pandemic raised on the end of December 2019 has drastically affected different aspects of human wellbeing. The pandemic not only affected the health of individuals, but also negatively affected mental health and social interaction. This review illustrates: a) scientific findings related to the impact of the COVID-19 pandemic on the reproductive process, considering gender, hormonal balance, gonad functions, pregnancy, and ART, b) the sociosexual dimension of COVID-19 disease and precautions that should be taken to avoid infection via sexual transmission or vertical transmission, which may alleviate the fear associated with continuing normal social relationships and economic activities.

Sex preselection of sٍheep embryo by altering the minerals of maternal nutrition.

Several methods have been conducted for embryo sex preselection, which includes X- and Y- sperm separation, changing the pH of the female reproductive tract, time of mating before or after ovulation, and feeding formula, such as altering the presence of minerals in diet content before breeding may affect the embryo sex preselection ratio. In this study, three food formulas to feed female sheep were created with the cooperation of the Arabian Agricultural Services Company (Arasco). Ewes were fed with modified food formulas for one month before mating with males. The first group (A) (30 ewes), modified for male embryo gender preselection, were fed a diet with an increased percentage of the minerals Na+, K+, and P-. The second group (B) (30 ewes), modified for female sex preselection, were fed a diet with an increased percentage of the minerals Ca++ and Mg++. The third (control) group (C) (30 ewes) were fed the regular (Wafi) food formula. Our results showed no significant differences were in mean body weights between the three groups at the end of the feeding period. The results of different feeding formulas on mineral serum blood samples of ewes showed an increase in Na+, K+, and Cl- ions in the serum of group (A) compared to the other groups (B and C). The concentration of Na+ in the serum of group (A) was significantly (P < 0.05) higher than group (C). The concentration of Cl- ions in serum samples of ewes in group (A) was significantly higher than group (C) and group (B) (P < 0.05). The role of maternal feeding on embryo sex preselection shows that the pregnancy rate of animals in group (A) was 73.33%. Group (A) birthed 17 males and 5 females (77.27% and 22.72%, respectively). The pregnancy rate in group (B) was 70%. Group (B) birthed 6 males (27.27%) and 16 females (72.72%). Finally, the control group (C) had a pregnancy rate of 76.66%. They birthed 13 males (54.41%) and 11 females (44.83%). The results of our study confirm that altering the percentage of minerals in the maternal diet plays a role in sex preselection in sheep, which agree with other mammalian studies in rats and mice. Thus, the result of this study can help farmers to manage their breeding. We recommend that more studies on the relationship between minerals in the diet should be conducted for other spices and human sex preselection.

Effects of Varying Dietary DL-Methionine Levels on Productive and Reproductive Performance, Egg Quality, and Blood Biochemical Parameters of Quail Breeders.

The present study was carried out to study the effects of varying dietary DL-methionine (0, 0.5, 1.5, 2.5, and 3.5 g/kg) levels on the productive and reproductive performance, egg quality and blood biochemical parameters of quail breeders. In total, 150 mature Japanese quails at eight weeks of age were randomly allotted to five groups of 30 for each group. Each group included five replicates, each of six quails (four females and two males). The results showed that egg number, egg weight and egg mass were higher (p < 0.05) with the addition of all DL-methionine levels than that of the control group. Quails from the control group had a lower feed intake (p < 0.001) and a worse feed conversion ratio (FCR) than those from the DL-methionine-treated groups. Supplementation of DL-methionine up to 2.5 g/kg in quail diets increased fertility and hatchability percentages. Birds fed DL-methionine at 1.5 g/kg had the best egg production indices, better FCR and the highest values of fertility and hatchability. Egg weight, yolk %, Haugh unit, egg shape index and unit surface shell weight (USSW) were increased and eggshell % was decreased in quail supplemented with DL-methionine levels compared with the control quail (p < 0.05). Dietary DL-methionine levels did not affect (p > 0.05) the hemoglobin (Hb), red blood cells (RBCs), white blood cells (WBCs) and packed cell volume (PCV) of quails. DL-methionine levels (0.5 and 2.5 g/kg) augmented lymphocytes and basophile (p < 0.05). Low DL-methionine levels (0.5 or 1.5 g/kg) improved liver enzymes and kidney functions. Dietary DL-methionine levels (except 3.5 g/kg) declined serum lactate dehydrogenase (LDH) and decreased lipid profile parameters (except high-density lipoprotein-HDL). Supplementation of DL-methionine at 0.5 and 1.5 g/kg increased immunoglobulin (IgG, IgM and IgA) and superoxide dismutase (SOD), total antioxidant capacity (TAC), catalase (CAT) and reduced glutathione (GSH) (p < 0.001) compared with the control. In conclusion, dietary supplementation of DL-methionine (1.5 g/kg) can enhance the reproductive performance and egg quality of quail breeders. DL-methionine use at levels of 0.5 or 1.5 g/kg improved the liver and kidney functions, lipid profile, immunity and antioxidant parameters of Japanese quail.

The in vitro development of cloned sheep embryos treated with Scriptaid and Trichostatin (A).

Although, it has been success in the generation of animal clones from somatic cells in various animal species, the information related to nuclear reprogramming of cloned embryos is found to be limited. This study aims to compares the effect of both Scriptaid (SCR) and Trichostatin (A) treatments in improving cloning efficiency, and embryos developmental rate of cloned sheep embryos in vitro. Three groups were formed, i.e., one SCR group, second TSA group, with both treatment concentrations of 5 nM, 50 nM, and 500 nM, respectively, and third were control group with 0 nM. Methods: Ovaries of slaughtered sheep were collected and oocytes were recovered from antral follicles using aspiration method and in vitro maturation of oocytes were done. Then zona dissecting with micropipettes and oocyte enucleation were carried out under the micromanipulator. Later nuclear transfer, cell fusion and activation were done via cell fusion machine. Finally the embryo cultured in incubating chamber at the CO2 incubator up to 9 days. The result: In general the results showed that when the concentration increases the cleavage rate increased. The cleavage rates of the SCNT embryos treated with SCR at different concentrations are closely related to cleavage rate of embryos treated with TSA at same concentration; such as 39.47% for 500 nM TSA, 38.09% for 500 nM SCR; 18.6% for 50 nM TSA, 19.17% for 50 nM SCR, and 22.64% for 5 nM TSA, 17.18% for 5 nM SCR. As for the control group, the cleavage rate of the SCNT embryos cleavage ratewere27.47%., 30% and 30.85% respectively for bothtreatments. While there is a significant difference in TSA treatments at an eight-cell stage at the concentration (5 and 50 nM TSA) compared to the all other cleavage cell stages of (500 nM TSA and control). Also their were a differences between (50 nM of TSA) compared to the (50 nM SCR). Also there were a significant differences between the 16 cell stage at the (500 nM TSA) compared to other treatment (5 nM, 50 nM TSA and control). Regarding the SCR there were a significant difference at 8 cell stage between (5 nM SCR), compared to the other treatment (50 nM, 500 nM SCR and control). Also there were a significant difference at 16 cell stage between (50 nM, and 500 nM SCR), compared to the other treatment (5 nM SCR and control). While in the development of the embryos reach to blastocyst stage the SCR and the control group show a higher rate, in compered to TSA that did not show any development to blastocyst stage. The total SCR treatment showed (3/41 = 7.31%), and the total control showed (4/89 = 4.49%) blastula stage. It concludes that SCR improve the final development blastula stage compared to the TSA treatments that did not improved embryos reach to final developmental blastula stages may be due to spices differences or to the toxicity of TSA, especially at higher concentrations.

The Effect of Adding Different Levels of Curcumin and Its Nanoparticles to Extender on Post-Thaw Quality of Cryopreserved Rabbit Sperm.

The cryopreservation process adversely affects sperm function and quality traits, causing some changes at biochemical and structural levels, due to mechanical, thermal, osmotic, and oxidative damage. Supplementation with curcumin nanoparticles could prevent and even revert this effect and could enhance the post/thawed sperm quality in the rabbit. The study amid to explore the effect of curcumin (CU) and curcumin nanoparticles (CUNPs) supplementation in semen extender on post/thawed rabbit sperm quality. Twelve fertile, healthy rabbit bucks were included, and the ejaculates were collected using artificial vaginas. Rabbit pooled semen was cryopreserved in tris-yolk fructose (TYF) extender without any supplement (control group) or extender supplemented with CU at levels of 0.5, 1 or 1.5 µg/mL (CU0.5, CU1.0, and CU1.5, respectively) or CUNPs at levels of 0.5, 1, 1.5 (CUNPs0.5, CUNPs1.0, and CUNPs1.5, respectively) and was packed in straws (0.25 mL) and stored in liquid nitrogen (-196 °C). Results revealed that CUNPs1.5 had a positive influence (p < 0.05) on post-thawing sperm progressive motility, viability, and membrane integrity as compared with the other groups. Percentages of dead sperm, abnormalities, early apoptotic, apoptotic, and necrotic sperm cells reduced (p < 0.05) in CUNPs1.5 as compared to other treatments. Using 1.5 µg/mL of CUNPs significantly improved total antioxidant capacity (TAC), GPx, while MDA and POC reduced (p < 0.05) in CU1.5 in comparison with other groups. SOD values were enhanced (p < 0.05) in CUNPs1.0 and CUNPs1.5 in relation with other treatments. Conclusively, the addition of curcumin and its nanoparticles to the extender can improve the post-thawed quality of rabbit sperm via redox signaling and reduce the apoptosis process.

The relationship between cytokine gene polymorphism and unexplained recurrent spontaneous abortion in Saudi females.

OBJECTIVE: To investigate the relationships between unexplained recurrent spontaneous abortion (RSA) and single nucleotide polymorphisms tumor necrosis factor-alpha (TNF-alpha) (-238 G/A, -308 G/A), interleukin (IL)-6 (-634 G/C) and IL-10 (-592 C/A) in the promoter region of 3 different interleukin (TNF-alpha, IL-6, and IL-10) genes. METHODS: The study group comprised 65 women (mean age: 34.1+/-6.2; range: 15-45 years) with unexplained RSA, consecutively referred to the Recurrent Abortion Clinic, King Khaled University Hospital, Riyadh, Kingdom of Saudi Arabia from January 2010 to January 2011. The control group consisted of 65 females with at least 2 successful pregnancies and no history of abortion. Blood samples were drawn and deoxyribonucleic acid (DNA) was extracted using Puregene DNA purification kit. Utilizing polymerase chain reaction, the promoter region was amplified and sequenced on an Applied Biosystems Integrated sequencer to study the polymorphic sites of interest. All polymorphisms were identified in the case and control samples. RESULTS: A significant association was identified only between the -308 G/A polymorphism in the TNF-alpha gene promoter and the occurrence of unexplained RSA, and there was no significant association with other positions. CONCLUSION: The TNF-alpha gene polymorphism at position -308 could be a genetic predisposing factor for unexplained RSA.