Genital Tract Affections in the Female Buffalo

A healthy genital tract is a prerequisite for a buffalo to maintain its ability to reproduce and produce milk. Any abnormality in the tubular genital tract would hinder or lower the passage of gametes and embryos and their establishment in the uterus resulting in lower fertility in buffaloes which inherently have low fertility [1] and which is also largely dictated by environmental and photoperiodic cues. Numerous abattoir surveys of buffalo genitalia have been conducted to investigate macroscopic and microscopic abnormalities [2-9]. A wide variation has been recorded in the incidence of abnormalities found over different geographical locations. These findings are affected by various factors, such as incidence of diseases, amount of veterinary supervision and critical appraisal of abnormalities by the person carrying out the survey [10]. At organized farms, reproductive problems were the biggest cause (38.62%) of culling of Murrah [11] and Bulgarian Murrah (41.0%) buffaloes [12].

Genital affections appear to be an important reason for culling infertile buffaloes. A recent study on abattoir derived genital organs [13] showed the greatest number of affections in the cervix (51.91%) followed by the uterus (29.01%), ovaries and other structures. Two thirds of culled buffaloes had either a single or multiple gross abnormalities and acquired cervical affections were observed to be the most frequent in half of the culled buffaloes. Such studies are suggestive of sequelae of multiple cervical insults / injuries while handling the female genitalia during parturition or artificial insemination. Descriptions on the different affections of the female genital tract of buffaloes have poor clinical orientation and little adaptability because of the ease with which animals can be slaughtered. In recent years, clinical descriptions on the various affections of the bubaline genital passage have been conducted and many more new methodologies of diagnosis and therapy are being adopted.

1. Genital Affections

1.1. Vaginal and Vulvar Affections

Vaginal affections include vaginitis and lacerations; congenital problems include imperforate hymen, tissue bands, double vagina and acquired abnormalities include stenosis, retention cysts, uro-vagina and tumors [14,15]. Unusual cases of muco-vagina have been reported [16].

Vulvar affections include vulvitis, granular vulvo-vaginitis and vulvar lacerations [17,18]. The incidence of vulvo-vaginal affections in abattoir studies was 4.9 % [13,15] whereas in clinical studies the incidence of vaginitis was 0.71 % [19].

1.1.1. Etiology

Facultative microorganisms like E. coli, Klebsiella, Micrococcus sp, and Staphylococcus aureus are normal inhabitants of the bubaline vagina [20,21]. The vaginal microflora changes depending upon the stage of reproduction such as estrus, pregnancy and parturition [21,22]. Buffaloes with inactive ovaries show a higher rate of bacterial presence in the vagina [22]. In a recent report, Chlamydia was isolated from vaginal swabs from buffaloes with or without signs of reproductive disease [23]. Vaginitis may also result from the improper use of instruments, wallowing and poor hygiene [9]. The etiology of acquired vaginal problems like retention cysts, fibrous bands and tumors appear to be due to trauma, especially during parturition. Granular vulvo-vaginitis is caused by herpesvirus in the buffalo [24]. Vaginal and vulvar lacerations are frequently encountered during dystocia handling.

1.1.2. Clinical Diagnosis

The diagnosis of vaginal and vulvar affections is easy. A vaginoscopic examination (Fig. 1) would reveal inflammatory lesions, tissue bands or growths in the vagina and lesions of the vulva are visible externally. Biopsy specimens of tumors can be obtained from the vagina for histopathology. Uro-vagina can also be visualized. Vaginal / vulvar stenosis can result in painful mating. Pustular vulvovaginitis is characterized by vulvo-vaginitis in female buffaloes with discharge of pus and painful lesions. The disease may be prevalent for 5-10 days. Swabs for diagnosis of pustular vulvovaginitis should be carefully collected and submitted to competent laboratories [24]. Buffaloes with vaginitis can develop vaginal prolapse [25] and can attract males without being in estrus.

Figure 1. Vaginoscopic examination to evaluate the vagina and external os of the cervix in a buffalo.

1.1.3. Therapeutic Considerations

Vaginitis can impair fertility in naturally mated buffaloes. Some of the other congenital defects described earlier may have similar effects on fertility. Vaginal infections can be treated by careful vaginal douches with antibiotics, antiseptics or application of emollient creams. Surgical correction of congenital defects is not possible under most clinical settings. Culling such animals should be suggested. Removing these animals from the herd will probably prevent the genetic transmission of the congenital defect. Imperforate hymen or vaginal tissue bands can be surgically excised. Surgical correction of uro-vagina necessitates specialized instruments and expertise that preclude their frequent use in buffaloes.

1.2. Cervical Affections

Cervical affections in buffaloes include cervicitis, congenital defects, kinked cervix, cervical cysts, tumors, cervical lacerations and cervical fibrosis [14,17,26-30]. Of the various affections of the cervix, cervicitis is considered the most common. The clinical incidence of cervicitis is known to vary from 0.5 to 10% [31]. In a clinical study of 16,538 buffaloes, the incidence of cervicitis was low (2.09%) [19]. In other clinical surveys, the incidence of cervicitis was 0.36% [32], 0.5-2% [33] and 11.60% [34]. Similarly, chronic cervicitis was observed in 9.6% of 760 buffalo uteri obtained from abattoir [15]. Rao and Raja [35] recorded an incidence of 0.52% cervical affections in buffalo genitalia obtained from an abattoir. Congenital abnormalities of cervix recorded for the buffalo include double cervix (Fig. 2 and Fig. 3) and cervical agenesis [9]. Kinked and cervices with fibrosis are rarely encountered in buffaloes and the incidence of other affections like tumors and cysts are low, fibroma is the only tumor recorded for the buffalo [36].

Figure 2. The reproductive tract of a buffalo heifer showing two cervical canals (Photo Courtesy Prof. Azawi O.I., Department of Surgery and Theriogenology, College of Veterinary Medicine, University of Mosul, Mosul, Iraq).

Figure 3. The reproductive tract of a buffalo heifer cut open showing the two totally separate cervical canals (Photo Courtesy Prof. Azawi O.I., Department of Surgery and Theriogenology, College of Veterinary Medicine, University of Mosul, Mosul, Iraq).

1.2.1. Etiology

Cervicitis commonly develops subsequent to an ascending or descending infection from the genital tract [14] and is precipitated by the same factors that cause endometritis, vaginitis or both. Faulty insemination procedures and obstetric trauma may culminate in cervical infections. If left untreated, chronic cervicitis may develop with fibrosis and permanent enlargement of the cervix. Cervical cysts may develop due to constant pressure from irritating fluids or trauma [30,37]. Fibrous tissue bands in the cervix that attach to the vagina may have traumatic origins. The etiology of cervical tumors in the buffalo continues to be poorly understood.

1.2.2. Clinical Diagnosis

Trans-rectal palpations and vaginoscopy are the most frequently used procedures for diagnosis of cervical affections. The rectal palpation of a grossly enlarged cervix is a sufficient clue for diagnosis of cervical inflammation or hypertrophy. When visualized through a vaginal speculum, the inflamed cervix appears to be swollen with the mucosa appearing cherry red to dark purple in color and the external folds are often prolapsed [14]. In chronic cervicitis, a hard fibrous mass (the inflamed cervix) can be readily palpated by trans-rectal palpation and visualized by speculum examination or evaluated by ultrasonography [29,38]. Other congenital and acquired affections like cysts, tumors, tissue bands or occlusions can also be visualized by vaginoscopy. Tissue specimens can be collected for diagnostic evaluation of tumors through biopsy instruments and preserved in formol saline to be sent to the diagnostic laboratory. A cervical tumor may sometimes be hanging out of the vulvar lips. Anatomic defects or congenital malformations of the cervix can sometimes be palpated although they have mostly been detected in abattoir material like the two cervical canals shown in Fig. 2 and Fig. 3.

1.2.3. Therapeutic Considerations

Cervicitis should be treated promptly as persistence of infection tends to render the condition chronic with development of fibrous tissue, which then becomes irresponsive to treatment. As the condition coexists with vaginitis and endometritis, it is better to treat the entire genital tract using antibiotic infusions, both intra-uterine and intramuscular [39]. Cervical tears at parturition need prompt surgical repair when they are large. Application of emollient creams and smoothing agents are often helpful in treating the condition. Cervical fibrosis and kinks developing in the cervix due to trauma or constant infection pose difficulties in the passage of insemination guns during artificial insemination. β2 adrenergic drugs like isoxsuprine hydrochloride 60-100 mg administered IV 15-20 minutes before insemination often relaxes the cervix and facilitate the passage of the gun [40]. Surgical excision of tumors of the cervix is possible under epidural anesthesia. The administration of 10-20 mg IM of xylazine may be helpful in retracting the cervix out of the vulvar lips for surgery. The affected tumor masses are removed after proper ligation. Sufficient postoperative application of emollient antibiotic cream is suggested for 10-15 days for proper recovery.

1.3. Uterine Affections

The affections of the uterus include congenital anatomic defects [9], infections involving the different layers of the uterus (metritis, endometritis, perimetritis) [9], functional aberrations of the uterine mucosa (hydrometra, mucometra, pyometra), uterine adhesions with the surrounding structures [10] and uterine cysts and tumors. Uterine infections have been reported as the major cause of infertility in buffaloes [14]. A healthy uterus is the most vital component for the implantation and development of the embryo and fetus. The presence of bacteria in the uterus causes inflammation and lesions in the endometrium and delays uterine involution, perhaps influences embryo survival [41]. In addition, uterine inflammation compromises the proper function of the uterus, which could lead to infertility [42].

Anatomic defects of the bubaline uterus described from abattoir studies include uterus didelphus [43, 9], uterus unicornis and aplasia of the uterine horns [44,45]. Uterine tumors reported for buffaloes such as fibroma, are mostly benign [15] with its incidence being below 1 % [46]. Para-uterine and uterine cysts have been observed in abattoir genitalia [45].

Figure 4.1. A buffalo with vaginal prolapse.

Figure 4.2. A buffalo with vaginal prolapse.

Figure 4.3. A buffalo with dystocia. One leg and the head of the fetus are protruding through the vulvar lips.

Uterine infections involving different layers of uterus have been classified as metritis, endometritis, perimetritis and parametritis. The infections seem to be common during the postpartum period, although they may occur at other times as well [6]. The incidence of uterine infections in buffaloes depends upon the hygiene at calving. The risk factors for development of infection are prolapse (Fig. 4.1 and Fig. 4.2), dystocia (Fig. 4.3), retained fetal membranes [9] and metabolic disturbances during the post-partum period. The incidence of dystocia in buffaloes is known to be less than 10% [9,47-49] whereas that of retained placenta varies between 2.7 to 9.6% [12,50-53]. The incidence of endometritis has been widely studied from both abattoir material and clinical cases (Table 1). In general, a higher incidence has been observed for buffaloes compared to cattle [9]. The same definitions for uterine infection are used in cattle [41] and in buffaloes [9] and therefore, metritis and toxic puerperal metritis are considered acute inflammatory conditions of the uterus with or without systemic illness with discharge of purulent material during the first two weeks postpartum. Clinical endometritis and pyometra are considered closely related entities occurring 3-4 weeks post-partum where the latter occurs after the first postpartum ovulation. Purulent or mucopurulent discharge is characteristic in clinical endometritis whereas pus increasingly retained in the uterus and persistent corpus luteum is characteristic in pyometra. Based on uterine cytology, if endometritis occurs 8 to 15 weeks postpartum, it is considered as subclinical endometritis [9,54]. Repeat breeding is the major clinical consequence in buffaloes with clinical and subclinical endometritis. Endometritis has been recorded in buffalo heifers [6].

1.3.1. Etiology

Most postpartum uterine infections are due to poor parturient hygiene, vaginal stimulation for milk letdown, immune functions of the uterus and probably wallowing [9]. Microbes present in the environment may frequently infect the uterus iatrogenically during difficult births, retained fetal membranes and vaginal or uterine prolapse. The cause of uterine infections at times other than parturition could be due to breeding with infected bulls, poor hygiene during insemination or infection by hematogenous route. The cause of uterine abnormalities such as cysts or tumors are difficult to trace, although the constant pressure of irritating fluids, or tissue debris could be a possible cause. Pyometra is the accumulation of purulent material within the uterine lumen in the presence of a persistent corpus luteum and a closed cervix and is often closely associated with endometritis. The accumulated pus prevents further luteolysis. Post-service pyometra due to Trichomonas fetus infection has not been reported in buffaloes. Mucometra is the sequel of ovarian cysts or endometrial malfunctioning. Higher albumin and cholesterol in uterine fluids was recorded in mucometra affected buffaloes [55].

1.3.2. Clinical Diagnosis

The clinical diagnosis for uterine affections is first described for the different uterine infections and subsequently for other affections.

Figure 5. A buffalo with the retained fetal membranes hanging out of the vulvar lips.

1.3.2. (A) Metritis

Metritis is usually the result of retained fetal membranes (Fig. 5) and dystocia [56]. Many authors find that prevalence of metritis in buffalo ranges from 2.7% to 12.5% [12,51-53,57-58]. Bacteriological studies have shown that most prevalent bacteria in the vagina of metritis-afflicted buffaloes are Escherichia coli, Klebsiela pneummoniae, Lactobacillus acidophilus and Pseudomonas aeruginosa while the most commonly found bacteria in the uterus are E. coli and Archanobacterium pyogens [57]. Chronic metritis is found in a large percentage of buffaloes (58%) and most of them are severe cases. The proportion of sub-acute and acute metritis is 28% and 14% respectively [57].

Toxic puerperal metritis is an acute systemic illness due to a bacterial infection of the uterus, usually within 10 days of parturition. Affected buffaloes show a fetid, red, brown, watery discharge, reduced milk yield, dullness, anorexia and dehydration. [59]. The affected buffaloes have significantly higher creatine, kinase and aspartate aminotransferase in their blood. Clinical diagnosis can be based on these clinical signs and laboratory analysis. Affected buffaloes may be prone to developing metabolic disorders and frequently develop toxemia.

An abnormal large uterus and a purulent uterine discharge detectable in the vagina are signs of metritis, although these buffaloes are not systemically ill [9]. Diagnosis can be reached by trans-rectal palpation of the enlarged uterus, delayed involution is observed. Generally, the uterus of a parturient buffalo is palpable within the pelvic cavity after 10-12 days; but in the case of a buffalo with metritis and endometritis, the uterus is located in the abdominal cavity sometimes with a thick wall. Ultrasonographic evaluation of the postpartum uterus and its dimensions can help in monitoring uterine involution and the presence of excessive fluids in the uterus (Fig. 6 and Fig.7) whereas a vaginoscopic examination is useful in visualizing the presence of pus or mucus in the vagina. Lochial discharges during normal parturition should not be confused with metritis and can be differentiated by their scanty amount and absence of foul smell. The pathogenesis of uterine infections and the uterine defense mechanisms in buffaloes have been recently reviewed [9].

Figure 6. Ultrasonogram of a bubaline uterus after abortion showing fluid accumulation.

Figure 7. Ultrasonogram of a buffalo suffering from metritis. Echogenic material is visible within the uterine lumen.

1.3.2. (B) Endometritis

Endometritis is a very common disease in buffaloes and it is observed mostly during the postpartum period. The incidence of endometritis in water buffaloes (Table 1) is usually less than 20% [60-65]. A higher prevalence of endometritis was observed in some studies in India, Egypt and Iraq [66-68]. The difference in the prevalence of this condition can be due to the difference in breeds, nutrition and management [61]. The traditional practice also has an influence on the incidence of endometritis. Some farmers insert foreign material into the vagina of buffaloes to stimulate milk let down during the postpartum period. This habit is a predisposing factor for uterine infections in water buffaloes [57].

Figure 8.1. A buffalo with endometritis with mucoid vaginal discharge.

Figure 8.2. A buffalo with endometritis with mucopurulent vaginal discharge.

Clinical endometritis is characterized by the presence of a purulent or mucopurulent discharge detectable in the vagina usually 4-8 weeks postpartum although such discharges can appear without parturition in endometritis affected buffaloes or buffalo heifers. Diagnosis is based on the findings of discharges (Fig. 8.1 and Fig. 8.2), vaginoscopy and ultrasonographic appearance of uterine fluid with flakes of pus. Endometrial thickening and fluid accumulation can also be visualized by ultrasonography (Fig. 9.1 and Fig. 9.2).

The use of instruments such as Metricheck to detect endometritis in cows effectively can also be used in buffaloes. Another kind of endometritis is the subclinical endometritis, which is usually diagnosed by identification of PMNs in uterine cytology smears (Fig. 10.1and Fig. 10.2) obtained by the cytobrush or low volume of uterine flush [9].

Figure 9.1. Ultrasonogram of a buffalo with endometritis showing endometrial thickening.

Figure 9.2. Ultrasonogram from a buffalo with endometritis showing fluid accumulation in the uterus.

Figure 10.1. Uterine cytology smear from a buffalo with subclinical endometritis showing several PMNs (Photo Courtesy Md. Zakiuddin, Department of Veterinary Gynaecology & Obstetrics, Veterinary College, Bidar, India).

Figure 10.2. Uterine cytology smear from the same buffalo with subclinical endometritis showing reduction in the number of PMNs subsequent to therapy (Photo Courtesy Md. Zakiuddin, Department of Veterinary Gynaecology & Obstetrics, Veterinary College, Bidar, India).

Animals affected with subclinical endometritis do not show any pathologic processes in the vaginal discharge. A clinical sign of subclinical endometritis is repeat breeding in the buffalo. Ultrasonographic evaluation for sub-clinical endometritis has a low sensitivity and specificity. The white side test (Fig. 11) has been described for the diagnosis of uterine infections in the buffalo [80] and involves boiling cervico-vaginal mucus with 10% NaOH. Clumping of the mucus and development of yellow color indicates that the animal is suffering from uterine infection.

Figure 11. Bubaline white side test for diagnosis of sub-clinical metritis (From left: mild (light yellow), moderate (yellow), severe metritis (deep yellow) and negative (no colour change).

The pathological changes that are observed in buffaloes affected with uterine infections have been a focus of many studies on abattoir derived uteri [2,10,15,79,81-84] and a few clinical studies [9,85]. There are varying degrees of inflammatory changes with infiltration of PMNs in acute conditions and lymphocytic infiltration, appearance of fibroblasts, macrophages and thickening of blood vessels in chronic uterine infections [9]. The endometrial glands show enlargement and a desquamated epithelium often visible in histopathology sections. There is peri-glandular and perivascular cellular infiltration. In chronic endometritis, peri-glandular fibrous tissue is visible and the serosa is thickened due to fibrosis (Fig. 12, Fig. 13, Fig. 14, Fig. 15, Fig. 16, Fig. 17, Fig. 18 and Fig. 19).

Figure 12. Microscopic picture showing cellular infiltration in the myometrium of a metritis affected buffalo (uterus obtained from abattoir material).

Figure 13.Microscopic picture showing cystic enlarged endometrial gland in an endometritis affected buffalo (H&E stained at 40X magnification).

Figure 14. Microscopic picture showing desquamated epithelium in an endometritis affected buffalo (H&E stained at 200 X magnification).

Figure 15. Microscopic picture showing peri-glandular cellular infiltration in an endometritis affected buffalo (H&E stained at 100 X magnification).

Figure 16. Microscopic picture of an endometritis affected bubaline uterus showing peri-vascular and peri-glandular infiltration (H&E stained at 200 X magnification).

Figure 17. Microscopic picture showing green colored fibrous bands around endometrial glands in chronic endometritis in a buffalo (Massons trichome at 200 X magnification).

Figure 18. Microscopic picture showing periglandular fibrosis in a buffalo endometrium due to chronic endometritis (H&E stained at 100 X magnification).

Figure 19. Microscopic picture of the uterus of a buffalo showing thick serosa due to fibrous tissue (H&E stained at 200 X magnification).

A large number of microorganisms have been isolated from the uterus and cervico-vaginal mucus of buffaloes affected with uterine infections. These include Streptococci, Staphylococci, E. coli, Arcanobacter pyogenes, Micrococcus sp., Bacillus sp., Pseudomonas aeruginosa and Mycobacteria and Klebsiella [9,79,86-89]. Often these organisms are present collectively along with anaerobic bacteria like Fusiformis necrophorus and Bacteroides sp. [9,90]. Fungi like Aspergillus fumigatus have rarely been isolated from bubaline uteri [90]. Arcanobacter pyogenes is considered the only organism associated with severe endometrial lesions in affected buffaloes [9].

1.3.2. (C) Perimetritis and Parametritis

These conditions are rarely noted in buffaloes and develop secondary to pyometra and metritis. There are varying amounts of adhesions between the uterus and the broad ligaments and other pelvic and abdominal organs. These conditions involve inflammation of the serosa and uterine ligaments and other pelvic and abdominal organs. The diagnosis is somewhat difficult by trans-rectal examination of genitalia. The incidence of parametritis and perimetritis was 0.7% and 1.5% in buffaloes [43,115]. The prognosis is usually grave and affected animals may become sterile. Perimetrial cysts are rarely observed in abattoir-derived genitals (Fig. 20).

Figure 20. Perimetrial cysts observed in abattoir-derived buffalo genitalia.

1.3.2. (D) Pyometra and Mucometra

These conditions are characterized by the accumulation of pus (Fig. 21) or fluid within the endometrial cavity, persistence of a corpus luteum and suspension of the estrous cycle. The incidence of pyometra has been reported to vary from 0.25 to 9.25% in buffaloes [43,71,91]. The incidence of mucometra was 1.78% in one of the study [10]. The trans-rectal palpation of genitalia and visual inspection of the vulva for purulent exudates are common methods used for diagnosis. Observation of purulent exudates with the aid of a vaginal speculum has been reported to be a useful tool for diagnosis. Trans-rectal palpation reveals a flaccid uterus, atonic and symmetrically enlarged uterine horns with or without crepitating sounds [92]. The uterine wall is thicker, doughier, has a less vibrant feel, no slipping of allanto-chorion is felt nor are the caruncles palpable. Real time ultrasonography (Fig. 22) can be used to demonstrate uterine changes associated with postpartum infections by observing echogenic particles with anechogenic fluid (Fig. 7) and thickened uterine wall [93]. The samples of uterine fluid should be cultured in both aerobic and anaerobic environments. Mucometra / hydrometra can be similarly diagnosed by the accumulation of fluid in both uterine horns (Fig. 23) with a thin uterine wall and the ultrasonographic appearance of fluid (Fig. 24) without cotyledons or fetal structures [29,38], which differentiates it from normal pregnancy and pyometra.

Figure 21. A bubaline uterus with pyometra. (Photo Courtesy Prof. Azawi O.I., , Department of Surgery and Theriogenology, College of Veterinary Medicine, University of Mosul, Mosul, Iraq).

Figure 22. A sonogram of a bubaline uterus with pyometra. Echogenic material is visible within the lumen of the uterus.

Figure 23. A bubaline uterus with hydrometra with bilateral enlargement and fluid filled cavity.

Figure 24. Ultrasonogram of a buffalo uterus with anechoic fluid accumulation without the presence of a fetus or, cotyledons, characteristic of hydrometra/mucometra.

1.3.2. (E) Uterine Cysts and Tumors

Uterine cysts and tumors can be diagnosed by ultrasonography when the size is sufficiently large to be visible by this approach. Hysteroscopy can help in evaluating the texture and morphology of the bubaline uterus (Fig. 25 and Fig. 26), however, only a few reports are currently available [129]. Laparoscopic visualization of the genital structures has been documented for the buffalo [94], but the invasive nature of the technique precludes its frequent use in buffalo. Trans-rectal palpation can detect large-sized growths or anatomical defects of the uterus although such defects have mostly been reported from abattoir material [15], probably because such buffaloes seemed to be infertile and were sent to slaughter. Trans-rectal ultrasonography can aid in the diagnosis of uterine cysts and tumors when performed carefully and when the size of the cyst or tumor is large enough to be visualized by ultrasonography. Tumors often appear as heterogeneous masses with varying echogenicity when viewed with the ultrasound. Uterine cysts and tumors (Fig. 27.1 and Fig. 27.2 ) often hinder the normal progression of pregnancy. Cows with uterine adenocarcinoma present clinical evidence of bleeding at estrus and such signs are likely to be seen in buffaloes although they are not reported in this species.

Figure 25. Hystreroscopic view of a normal bubaline uterus.

Figure 26. Hemorrhages visible with hysteroscopy in two locations in a buffalo uterus.

Figure 27.1. A fibroma recovered from an abattoir derived buffalo uterus.

Figure 27.2. Histopathological section of the fibroma revealed green colored fibrous whorls (Massons Trichome 40 X magnification).

1.3.3. Therapeutic Considerations

A wide variety of therapeutic approaches has been mentioned for uterine infections in buffaloes and includes the administration of antibiotics, hormones, uterine motility stimulators and immunomodulators.

1.3.3. (A) Metritis

Arguments on the use/non-use of intrauterine antibiotics in the therapy of postpartum uterine infections continue, yet antibiotics are the most commonly used therapy. The acute nature of postpartum metritis and its harmful effects on milk production necessitates prompt therapy. Most antibiotics administered by intrauterine route are ineffective (first 10-12 days postpartum) when the condition occurs postpartum because of the presence of a large volume of fluid and tissue debris. Therefore, it is suggested to use antibiotics such as oxytetracycline [9], gentamicin, cephapirin or ceftiofur IM. Sufficient fluid replacement should be given in more severe cases with toxemia. Buffaloes with metritis are often anorectic and thus require the administration of liver tonics and antihistaminics. The administration of 250-450 mL of calcium borogluconate often helps in restoring uterine motility but such administration should be avoided if the animals are hyperthermic. High fever should be controlled by the administration of paracetamol, analgin or other appropriate drugs IM. Uterine clearance of purulent fetid material can be promoted by intrauterine administration of 500 mL of normal saline and IM administration of prostaglandins [95].

1.3.3. (B) Endometritis

A large number of antibiotics have been suggested to be administered intrauterine to treat clinical and subclinical endometritis in buffaloes (Table 2). It is suggested to administer antibiotics for 3-5 days or 6-8 h post-insemination in affected buffaloes. A combination of antibiotics against both aerobic and anaerobic microbes has been suggested [40]. The IM administration of 2 g of ceftiofur for 2-3 days has shown some promise in cows [96,97] and would probably work similarly in buffaloes. In chronic endometritis the presence of fungi is likely. The administration of anti-fungal products has been useful. Most studies have evaluated the success rates in terms of pregnancy rates subsequent to therapy [31,74,88], however, there is a lack of accuracy as to the effectiveness of the administration of antibiotics on the clearance of pathogens from the uterus. The pharmacokinetics of most antibiotics administered via the intrauterine route, are poorly understood. Thus, the dosage and frequency of administration are based on clinical usage. The antibiotic sensitivity pattern of microbes isolated from the cervico-vaginal mucus from endometritis-affected buffaloes had been the focus of many studies with diverse results [83,90,98,99]. The cervico-vaginal mucus may not be a true representative of uterine microbes as it passes through the vagina where facultative organisms are frequently located. Therefore, it is suggested to use specialized double barrel swab collection pipettes (Fig. 28) for collection of uterine samples and isolation of microbes. Moreover, the type of organisms may vary in number, virulence and resistance towards antibiotics at different locations. Antibiotic sensitivity against microbes in the cervico-vaginal mucus may not be an optimal approach for selecting the antibiotic to treat endometritis.

Figure 28. Double barrel uterine swab collection pipettes appropriate for bacterial culture and isolation.

It has been mentioned that, there is an increase in the free radicals in the uterine lumen of endometritis affected buffaloes [100], the intrauterine administration of antioxidants such as 200 mg of vitamin C, offers some promise in resolving endometritis and improving the uterine environment [40]. The IM or intrauterine administration of other antioxidants such as vitamin E may also be helpful in resolving endometritis in buffaloes [101].

The intrauterine use of immunomodulators is considered to evoke an influx of polymorphonuclear leukocytes inside the uterine lumen, increasing phagocytes’ activity and improving chemotaxis. The use of oyster glycogen and E. coli lipopolysaccharides (LPS) have shown some promise in resolving endometritis in buffaloes (Table 3).

The use of hormones like estrogens administered by the intrauterine or IM route has been disappointing in buffaloes. Estrogens had little effect on the clearance of uterine infection; however, reduced milk production in lactating buffaloes was noted [9]. The administration of prostaglandins in endometritis-affected buffaloes has shown some success [9,102] since prostaglandins increase uterine contractility and a return to estrus favors uterine clearance of microbes. Prostaglandins can be administered after 5-10 days of estrus in buffaloes at a dosage similar to the one used in cattle [40]. Being a pro-inflammatory cytokine, the administration of prostaglandins may rarely precipitate inflammation of limbs and lameness.

1.3.3. (C) Perimetritis and Parametritis

The therapy of these conditions is usually not possible. Laparoscopic detachment of the adhesions can be attempted but economic considerations and the possibility of reformation limit the use of such approaches. Prognosis of perimetritis in cows is extremely poor, most cows with this condition die within a week after diagnosis. Similar outcome is likely with affected buffaloes although not documented.

1.3.3. (D) Pyometra and Mucometra

Pyometra and mucometra in buffaloes can be successfully treated by the administration of prostaglandins. The pus/fluid is usually discharged within 48 h of prostaglandin administration. In the more severe cases, it is suggested to administer prostaglandin twice 7 days apart. In pyometra, uterine lavage with 1L of normal saline is suggested for 3-7 days to evacuate the uterus of tissue debris and purulent material. Daily trans-rectal uterine massage and manual backward pressure on the uterus often helps in the clearance of the purulent material. Buffaloes with mucometra often recover with a single administration of prostaglandins. It is often useful to supplement affected buffaloes with 10-15 g potassium iodide orally (daily for 5-7 days) or with SC administration of iodine solutions.

1.3.3. (E) Uterine Cysts and Tumors

Treatment of cysts and tumors involves surgical removal via laparotomy. However, affected buffaloes are seldom treated, rather, these animals are slaughtered.

1.4. Affections of the Oviduct

Healthy oviducts are essential for the fertilization of gametes and maintenance of the embryo until it is transported into the uterus. Affections of the oviducts include congenital defects, salpingitis, hydrosalpinx, pyosalpinx and adhesions; these are uncommon causes of infertility and sterility in buffaloes. Affections of the oviducts result in occlusion of the lumen preventing fertilization or creating an unfavorable environment for fertilization. A unilateral affection results in infertility, whereas a bilateral affection results in sterility. Affections of the oviducts have been diagnosed largely from abattoir studies. The overall incidence of affections of the oviduct varies from 10 to 29% [61,69,115-117], of which salpingitis is the most common [118]. A higher incidence of oviduct affections is reported in buffaloes compared to cattle [46,119,120]. The various affections of the oviduct revealed slight variations in different studies (Table 4). Blood accumulations in the oviduct and double congenital oviducts have rarely been recorded in buffaloes [117].

1.4.1. Etiology

Salpingitis is usually caused by infections in the uterus, cervix or vagina. Buffaloes with salpingitis may be sterile depending on the severity of the condition. Salpingitis develops due to upward infection from the uterus following abortion, retained fetal membranes, septic metritis and pyometra. Tuberculosis is also one of the causes for oviductal lesions. Uterine irrigation with strong antiseptic solutions may escape into the oviducts and cause inflammation. A descending infection from the peritoneum also cannot be ruled out.

Hydrosalpinx is usually the result of non-infectious inflammation and/or serous secretions and has been reported to be a congenital disease [124]. In contrast, Mastroianni [125] believed that the condition was the result of inflammation around the oviducts. Hydrosalpinx (Fig. 29) was also suggested to be the sequel of salpingitis [126]. A bacteriological study of hydrosalpinx suggests that there is no association between hydrosalpinx with the presence of bacteria in the uterus. By contrast, pyosalpinx has a correlation with the presence of bacteria in the lumen of the uterus in buffaloes. In hydrosalpinx, the most prevalent bacteria are Actinomyces bovis and Corynebacterium bovis while those in pyosalpinx are Archanobacterium pyogens and Corynebacterium kutscheri [117]. Pyosalpinx usually occurs because of a severe infection from the uterus and has been associated with adhesions of the mesosalpinx and mesovarium.

Adhesions of the oviducts have been the subject of many studies. The condition is perhaps the result of inflammation around the oviduct. Adhesions of the oviduct (Fig. 30) may develop to the surrounding bursa due to constant infection or following rough trans-rectal manipulations.

Figure 29. Bilateral hydrosalpinx in the genitalia of a buffalo (Photo Courtesy Prof. Azawi O.I., Department of Surgery and Theriogenology, College of Veterinary Medicine, University of Mosul, Mosul, Iraq).

Figure 30. Complete ovario-bursal adhesions in buffalo genitalia (Photo Courtesy Prof. Azawi O.I., Department of Surgery and Theriogenology, College of Veterinary Medicine, University of Mosul, Mosul, Iraq).

1.4.2. Clinical Diagnosis

Affections of the oviducts usually remain undetected. An expert ultrasound operator will be able to detect affections of the oviduct. Mild forms of salpingitis are difficult to diagnose and are usually detected after slaughter. In its severe form, the oviduct becomes thick, hard and swollen and it feels like a cord at trans-rectal palpation. When there is a large accumulation of fluid in the oviduct, it feels like a big cyst (hydrosalpinx) at palpation. Hydrosalpinx is characterized by the distension of the oviduct filled with amber colored fluid. The dilated oviducts may have a diameter of 30 mm [61]. Hyperplasia of mucosal epithelium is visible in histopathology (Fig. 31). Ultrasonography can aid in the diagnosis of affections of the oviduct. These affections frequently pose threats to the normal passage of spermatozoa in the female tract preventing fertilization. It is therefore important to evaluate the patency of the oviducts. The common approach to evaluate the patency of the oviduct is to use the passage of air or Phenolsulfonphthalein (PSP) dyes into the uterus using a Foley catheter fixed to the uterus. If the oviduct is open without any occlusions, the air or dye readily pass through the oviduct within 3-5 min and the dye may appear in the peritoneal cavity or in the urine [121,127,128].

Figure 31. Microscopic picture of an oviduct of a buffalo showing mucosal epithelial hyperplasia in the form of multilocular structure (H&E stained at 40 X magnification).

1.4.3. Therapeutic Considerations

Treatment may not be useful and prevention is more advantageous. If infection in the uterus is detected and treated early then it prevents the spread of the disease into the oviducts. If both oviducts are involved, then the animal will become completely sterile and has to be culled. Overall prognosis in affections of the oviduct has usually been unfavorable except in mild infections. Parenteral as well as intrauterine infusion of broad-spectrum antibiotics may or may not be beneficial. Gentle handling of the genitalia, proper treatment, control of uterine infections and sexual rest can reduce the incidence of affections of the oviduct.