Theriogenology 84 (2015) 452–456
Contents lists available at ScienceDirect
Theriogenology journal homepage: www.theriojournal.com
A retrospective study on clinical findings of 7300 cases (2007–2014) of barren female dromedaries A. Ali a, b, *, R. Derar a, b, F. Al-Sobayil a, A. Al-Hawas a, K. Hassanein c a
Department of Veterinary Medicine, College of Agriculture and Veterinary Medicine, Qassim, Saudi Arabia Department of Theriogenology, Faculty of Veterinary Medicine, Assiut University, Assiut, Egypt c Department of Medical Microbiology and Immunology, Faculty of Medicine, Assiut University, Assiut, Egypt b
a r t i c l e i n f o
a b s t r a c t
Article history: Received 26 September 2014 Received in revised form 14 March 2015 Accepted 30 March 2015
The objective of this study was to investigate the clinical findings in barren female dromedaries examined for different complaints. Female camels were examined for repeat breeding with regular heat interval (RB-R, n ¼ 5444), refused mating (RM, n ¼ 1299), repeat breeding with long heat interval (RB-L, n ¼ 489), difficulties or bleeding during mating (DM, n ¼ 53), and for manifestation of male-like behavior (MB, n ¼ 15). The genital tracts of all females were evaluated using transrectal palpation, ultrasonography, and exploration of the vagina. Cervical swabs were obtained for bacteriologic examination. Clinical endometritis, ovarian hydrobursitis, and vaginal adhesions were the main clinical findings in the female camels examined for RB-R, RM, and RB-L, respectively. Parity affected the frequency of occurrence of these findings. The incidences of clinical endometritis, ovarian hydrobursitis, and vaginal adhesions in nullipara and multipara were 28% versus 32.3% (P ¼ 0.004), 37.1% versus 23.7% (P ¼ 0.001), and 5.7% versus 18.3% (P ¼ 0.001), respectively. Vaginal adhesions, persistent hymen, pelvic abscess, and vulvar atresia were the clinical findings in the female camels presented due to bleeding at mating or with a history of an incomplete intromission of the penis. The male-like behavior was associated with an enlargement of the clitoris and narrowing of the vulva and vagina. Trueperella pyogenes, Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumonia, Streptococcus zooepidemicus, and b-hemolytic Streptococcus were isolated from females presented for repeat breeding syndrome. In conclusion, clinical endometritis, ovarian hydrobursitis, and vaginal adhesions were the main clinical findings in barren female dromedaries. Parity affected the frequency of the clinical findings. Ó 2015 Elsevier Inc. All rights reserved.
Keywords: Female camel Infertility Endometritis Ovarian hydrobursitis Vaginal adhesion
1. Introduction Poor fertility is a major cause of economic loss. The reasons for low fertility are related to events before insemination or mating including inadequate oocyte and/ or follicular development, or after insemination or mating, due to failure of the fertilized ovum to undergo development [1–4]. The direct causes of low fertility are many This study was conducted at the Veterinary Teaching Hospital, Department of Veterinary Medicine, Faculty of Agriculture and Veterinary Medicine, Qassim University, Saudi Arabia. * Corresponding author. Tel.: þ966 16 3801480; fax: þ966 16 3801360. E-mail address: [email protected] (A. Ali). 0093-691X/$ – see front matter Ó 2015 Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.theriogenology.2015.03.039
and can be complex. Malnutrition, infections, congenital defects, management errors, and ovulatory or hormonal imbalances can all result in low fertility [5–8]. The problems of reproduction in the camel have not been as extensively investigated as in other animal species. The data collected regarding these problems have been derived mainly from information provided by camel owners and from slaughterhouse material. Very limited information has come from clinical and farm observations [5,7,9–11]. Examination of abattoir specimens has provided some information about possible diseases encountered in the reproductive tract, which include pyometra, bursal and ovarian adhesions, endometritis associated with a partially
A. Ali et al. / Theriogenology 84 (2015) 452–456
involuted uterus, and cystic ovarian degeneration [10]. In addition, more recent clinical studies have indicated ovarian hydrobursitis as being an important cause for infertility in dromedary camels [12–15]. Nawito [9] has recorded the bacteriologic findings in the uteri of 2075 dromedary camels of unknown history from Cairo abattoir. In 94 cases, clinical symptoms such as abscesses in the uterus, catarrhal endometritis, hemorrhagic endometritis, pyometra, and pyometra with macerated fetuses were found, along with the presence of Micrococcus pyogenes var. aureus as a prominent factor. Furthermore, the researcher was able to isolate Micrococcus pyogenes var. albus, beta-hemolytic streptococci, Escherichia coli, and Pseudomonas aeruginosa from the uteri of animals that had exhibited clinical symptoms. Studies on the etiology of reproductive disorders in female camels based on a large clinical database are needed to investigate the magnitude of each problem and to plan efficient strategies for their management. The objective of this study was to investigate the clinical findings in large number of barren female dromedaries examined for different complaints. 2. Materials and methods 2.1. Animals and management A total of 7300 female camels (Camelus dromedarius) aged between 5 and 18 years were examined for repeat breeding with regular heat interval (animals that show estrus at regular intervals, 12–14 days, but fail to conceive after three or more services, RB-R, n ¼ 5444), refused mating (a female camel that curls her tail dorsally when approached by a rutting male, RM, n ¼ 1299), repeat breeding with long heat interval (animals which refuse the male 2 weeks after mating as if they were pregnant; however, later on, >30 days, they return to estrus and accept the male, RB-L, n ¼ 489), difficulty or bleeding during mating (DM, n ¼ 53), and for manifestation of malelike behavior including vocalization, chasing estrus females, protrusion of the soft palate, and refusing to mate (MB, n ¼ 15). The average durations (standard deviation) of these reproductive disorders were 5 2.4, 17.5 12, 12 5.6, 7.8 2.5, and 9.5 1.8 months for RB-R, RM, RB-L, DM, and MB groups, respectively. The study was carried out during seven breeding seasons (from September 2007–February 2014). The examinations were conducted at the Veterinary Teaching Hospital of Qassim University, Saudi Arabia. Of the animals examined, 2263 (31%) were nullipara and 5037 (69%) were multipara. Body condition scoring, on a scale of 1 to 5 [16], ranged from 2.5 to 4. All examined females were generally healthy with no systemic illnesses. The majority of animals were left unconfined in open desert areas and fed mainly on alfalfa hay and barley concentrate. They were continuously exposed to fertile males during the breeding season. 2.2. Gynecologic examinations The reproductive tract of each animal was examined through standard transrectal palpation and by
453
ultrasonography using the Aloka SSD-500 model equipped with a 5-MHz linear-array transducer (Aloka Co., Ltd., Tokyo, Japan). The ovaries were examined for structure and size. The uterus was palpated for consistency, movability, and contents. Vaginal examination with a gloved hand was performed to estimate the patency of the vagina and cervix and to evaluate the nature of the vaginal discharges. Catarrhal (turbid mucus), mucopurulent (turbid mucus with flakes of pus), or purulent (profuse pus) vaginal discharges were regarded as signs of clinical endometritis [17–19]. According to the amount and viscidity of the vaginal discharges, degrees of clinical endometritis were categorized as (1) mild: females with small amount of mucopurulent vaginal discharge; (2) moderate: females with moderate mucopurulent discharge; and (3) severe: females with highly viscid and thick or pasty vaginal discharges. 2.3. Bacteriologic examinations A total of 45 female camels with repeat breeding syndrome (failure of conception in spite of mating with a fertile bull more than three times) were randomly selected for bacteriologic examination. Cervical swabs were obtained under aseptic conditions. All cervical samples were inoculated onto blood and MacConkey’s agar plates as described by Collee et al. [20]. The inoculated plates were incubated at 37 C for 48 hours. Gram-stained films were prepared from the growing colonies, and isolates were tested for motility. Gram-negative bacilli and gram-positive cocci were identified by means of API 20E and API 10S systems (Biomerieux SA, Montalien Vercica, France). A commercially available latex agglutination test was used for identification of Lancefield groups of Streptococci. 2.4. Statistical analysis The effect of parity on the frequency of different clinical findings in barren female dromedaries was evaluated using the chi-square test. GraphPad Prism program for Windows, version 300 (1999) was used for analysis. Significance was set at P < 0.05. 3. Results Clinical findings for the different forms of reproductive disorders are shown (Table 1). Clinical endometritis, ovarian hydrobursitis (fluid accumulation and encapsulation of the ovary by the ovarian bursa), and vaginal adhesions (occlusion of the vaginal passage) were the common clinical findings in female camels examined for RB-R, RM, and RB-L, respectively. Vaginal adhesions, persistent hymen, and vulvar atresia were the clinical findings in female camels presented due to bleeding at mating or with a history of an incomplete intromission of the penis (DM). Parity affected the frequency of occurrence of these clinical findings. Clinical endometritis and ovarian hydrobursitis were the common clinical findings in the barren multipara and nullipara, respectively (Table 2). Clinical endometritis was found in mild, moderate, and severe degrees in frequencies of 45%, 32%, and 23%, respectively. Vaginal discharges were observed after
454
A. Ali et al. / Theriogenology 84 (2015) 452–456
Table 1 Clinical findings in female camels examined for repeat breeding with regular heat interval (RB-R, n ¼ 5444), refused mating (RM, n ¼ 1299), repeat breeding with long heat interval (RB-L, n ¼ 489), and difficulty or bleeding at mating (DM, n ¼ 53). Clinical findings
RB-R
Clinical endometritis Ovarian hydrobursitis Vaginal adhesion Cervical stenosis Cervical adhesion Apparently normal genitalia Unovulatory follicles Inactive ovaries Hydrosalpinx Uterine adhesion Pneumovagina Persistent hymen Vulvar atresia Pelvic abscess Total
RM
RB-L
DM
Total
1890 (34.7) 1650 (30.3) 701 (12.9) 269 (4.9) 270 (5) 321 (5.9) 159 (2.9) 50 (0.9) 87 (1.6) 31 (0.6) 16 (0.3) d d
217 (16.7) 140 (10.8) 310 (23.9) 233 (17.9) 214 (16.5) 43 (3.3) 21 (1.6) 85 (6.5) 6 (0.5) 30 (2.3) d d d
152 (31.1) 245 (50.1) 6 (1.2) 28 (5.7) 5 (1) 20 (4.1) d d 33 (6.8) d d d d
5444
1299
489
d d 34 (64.2) d d d d d d d d 12 (22.6) 5 (9.4) 2 (3.8) 53
2259 (31) 2035 (27.9) 1051 (14.4) 530 (7.3) 489 (6.7) 384 (5.3) 180 (2.5) 135 (1.9) 126 (1.7) 61 (0.8) 16 (0.2) 12 (0.2) 5 (0.07) 2 (0.03) 7285
n (%)
exploration of the vagina but rarely during transrectal palpation. Additionally, uterine adhesions to the broad ligaments, rumen, or omentum were recorded. Bilateral and unilateral left and right ovarian hydrobursitis were observed at frequencies of 42%, 46%, and 12%, respectively. The affection was associated with clinical endometritis (21%), adhesions of the genital tract with the internal organs (15%), adhesion of the vagina and pyometra (8%), and distension of the fallopian tube (6%). The other cases had no apparent abnormalities in the genital tract other than the affected bursa. Vaginal adhesions were associated with the accumulation of hypoechogenic or hyperechogenic materials in the uterus in 92% of the cases. No uterine contents were found in the other cases. The vaginal adhesions were found at the caudal-, mid-, or cranial vagina in frequencies of 18%, 74%, and 8%, respectively. A hydrosalpinx could be palpated per rectum as a distended, fluctuated tortuous structure and detected ultrasonographically by the presence of hypoechogenic fluid within the fallopian tube. The affection was mostly unilateral (91.3%). Unovulatory follicles having either thick or thin walls were observed. Those with thick walls and internal trabeculae comprised 92.8%, and they were mostly multiple (79.6%). Persistent hymen was mostly (75%) accompanied by an accumulation of creamy fluids in the anterior vagina. Vulvar atresia characterized by complete occlusion of the vulva lips, with only a very small orifice found at the ventral commissure. One of these cases was accompanied by narrowing of the vagina. The male-like behavior was associated with an enlargement of the clitoris (12 of 15) and narrowing of the vulva and vagina (9 of 15). All these cases were nullipara and aged from 6 to 8 years. Seven cases had normal estrus behavior for about 8 to 12 months before showing masculinity. No ovarian cysts or tumors were detected in any of these cases. A total of 78 isolates were obtained from 39 of the 45 repeat breeding animals randomly selected for bacteriologic investigations; the other six animals had no bacterial isolates. Trueperella pyogenes (formerly named
Arcanobacterium pyogenes) and Staphylococcus aureus were the most commonly found isolates (Table 3). 4. Discussion According to the present data, clinical endometritis, ovarian hydrobursitis, and vaginal adhesions are the main clinical findings in the barren female dromedary camels. Generally, unhygienic measures during parturition, postpartum complications, and unsanitary gynecologic manipulation are contributing factors for uterine infections [7,21]. Venereal infection may also be involved. Pseudomonas aeruginosa, Klebsiella pneumonia, and Streptococcus zooepidemicus, which were reported in this study, are venereal diseases of concern in horses [22,23]. In agreement with the results of this study, Trueperella pyogenes, Staphylococcus aureus, Streptococcus zooepidemicus, Staphylococcus epidermidis, and b-hemolytic Streptococcus have been considered as common causes of endometritis in camels [7,9,24–26]. Endometritis causes
Table 2 Effect of parity on the incidence of different clinical findings in barren female dromedaries (n ¼ 7300). Clinical findings
Nullipara
Clinical endometritis Ovarian hydrobursitis Vaginal adhesion Cervical stenosis Cervical adhesion Apparently normal genitalia Unovulatory follicles Inactive ovaries Hydrosalpinx Uterine adhesion Pneumovagina Male behavior Persistent hymen Vulvar atresia Pelvic abscess Total
636 (28.1) 840 (37.1) 128 (5.7) 25 (1.1) 22 (1) 311 (13.7) 86 (3.8) 102 (4.5) 74 (3.3) 6 (0.3) 0 (0.0) 15 (0.7) 12 (0.5) 5 (0.2) 1 (0.04) 2263
Multipara
P value
1623 (32.2) 1195 (23.7) 923 (18.3) 505 (10) 467 (9.3) 73 (1.5) 94 (1.9) 33 (0.7) 52 (1) 55 (1.1) 16 (0.3) 0 (0.0) 0 (0.0) 0 (0.0) 1 (0.002) 5037
0.004 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.01 0.03 0.02 0.01 0.01 0.08 0.3
n (%)
A. Ali et al. / Theriogenology 84 (2015) 452–456 Table 3 Types and frequencies of cervical bacterial isolates in female dromedary camels presented for repeat breeding syndrome (n ¼ 45 animals). Bacterial isolates
Frequencies ()
Trueperella pyogenes Staphylococcus aureus Streptococcus pyogenes Corynebacterium diphtheroides Bacillus anthracoides Streptococcus zooepidemicus Staphylococcus epidermidis Enterococci Streptococcus b-hemolytica Coagulase –ve Staphylococcus Escherichia coli Proteus Pseudomonas aeruginosa Klebsiella pneumonia Total
14 13 9 8 7 7 5 5 2 2 2 2 1 1 78
infertility for the duration of the infection, and subfertility may persist, even after apparently successful treatment [27]. In addition, early postpartum endometritis has been known to have long-term influences on the steroid concentrations of the ovarian follicles long after the infection self-resolved. This is likely to affect oocyte quality and may partially explain the reduced conception rates and longer interval between calving and conception that are often associated with endometritis [28]. Clinically, most female camels diagnosed with endometritis were repeat breeders, but others refused mating, or accept mating after variable periods of anestrous. Clinical endometritis was detected more frequently in barren multiparous camels. This may be due to the fact that the uterus insulted by a variety of organisms during mating, clinical examination, parturition, and/or postpartum period [7]. In most females, infections are eliminated by natural uterine defense mechanisms. However, in a proportion of the females, these defense mechanisms fail partially or completely, allowing establishment of an infection [7]. Regimes consisted of intrauterine infusion of 100-mL acriflavin 0.1%, 100-mL lotagen 4%, or 300 mg/100-mL gentamicin sulfate accompanied with administration of PGF2a at infusion and hCG at mating have been found equally efficient in treating female camels affected with endometritis. The 90-day nonreturn and calving rates were 64% and 58.9%, 53.1% and 49.3%, and 53.3% and 42.5%, for the three regimes, respectively [29]. Ovarian hydrobursitis is a serious, long-standing, reproductive disorder in dromedary camels characterized by fluid accumulation and encapsulation of the ovary [12–14]. Many observations have suggested that ovarian hydrobursitis is initially an inflammatory process of the genital tract [13,14]. It seems that Chlamydophila abortus is responsible for the spreading of the ovarian hydrobursitis affection in dromedaries [14]. Chlamydophila could be transmitted by all methods of infection including the venereal route [30–32], which may explain the high incidence of infection among the barren nulliparous and multiparous camels. Surgical ablation of the affected bursa in unilaterally affected cases has been recommended because conception rates after these trials have been highly acceptable [12–14]; however, surgical ablation in bilaterally affected cases is not recommended. It is
455
possible to medically treat small-sized ovarian hydrobursitis (
Contents lists available at ScienceDirect
Theriogenology journal homepage: www.theriojournal.com
A retrospective study on clinical findings of 7300 cases (2007–2014) of barren female dromedaries A. Ali a, b, *, R. Derar a, b, F. Al-Sobayil a, A. Al-Hawas a, K. Hassanein c a
Department of Veterinary Medicine, College of Agriculture and Veterinary Medicine, Qassim, Saudi Arabia Department of Theriogenology, Faculty of Veterinary Medicine, Assiut University, Assiut, Egypt c Department of Medical Microbiology and Immunology, Faculty of Medicine, Assiut University, Assiut, Egypt b
a r t i c l e i n f o
a b s t r a c t
Article history: Received 26 September 2014 Received in revised form 14 March 2015 Accepted 30 March 2015
The objective of this study was to investigate the clinical findings in barren female dromedaries examined for different complaints. Female camels were examined for repeat breeding with regular heat interval (RB-R, n ¼ 5444), refused mating (RM, n ¼ 1299), repeat breeding with long heat interval (RB-L, n ¼ 489), difficulties or bleeding during mating (DM, n ¼ 53), and for manifestation of male-like behavior (MB, n ¼ 15). The genital tracts of all females were evaluated using transrectal palpation, ultrasonography, and exploration of the vagina. Cervical swabs were obtained for bacteriologic examination. Clinical endometritis, ovarian hydrobursitis, and vaginal adhesions were the main clinical findings in the female camels examined for RB-R, RM, and RB-L, respectively. Parity affected the frequency of occurrence of these findings. The incidences of clinical endometritis, ovarian hydrobursitis, and vaginal adhesions in nullipara and multipara were 28% versus 32.3% (P ¼ 0.004), 37.1% versus 23.7% (P ¼ 0.001), and 5.7% versus 18.3% (P ¼ 0.001), respectively. Vaginal adhesions, persistent hymen, pelvic abscess, and vulvar atresia were the clinical findings in the female camels presented due to bleeding at mating or with a history of an incomplete intromission of the penis. The male-like behavior was associated with an enlargement of the clitoris and narrowing of the vulva and vagina. Trueperella pyogenes, Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumonia, Streptococcus zooepidemicus, and b-hemolytic Streptococcus were isolated from females presented for repeat breeding syndrome. In conclusion, clinical endometritis, ovarian hydrobursitis, and vaginal adhesions were the main clinical findings in barren female dromedaries. Parity affected the frequency of the clinical findings. Ó 2015 Elsevier Inc. All rights reserved.
Keywords: Female camel Infertility Endometritis Ovarian hydrobursitis Vaginal adhesion
1. Introduction Poor fertility is a major cause of economic loss. The reasons for low fertility are related to events before insemination or mating including inadequate oocyte and/ or follicular development, or after insemination or mating, due to failure of the fertilized ovum to undergo development [1–4]. The direct causes of low fertility are many This study was conducted at the Veterinary Teaching Hospital, Department of Veterinary Medicine, Faculty of Agriculture and Veterinary Medicine, Qassim University, Saudi Arabia. * Corresponding author. Tel.: þ966 16 3801480; fax: þ966 16 3801360. E-mail address: [email protected] (A. Ali). 0093-691X/$ – see front matter Ó 2015 Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.theriogenology.2015.03.039
and can be complex. Malnutrition, infections, congenital defects, management errors, and ovulatory or hormonal imbalances can all result in low fertility [5–8]. The problems of reproduction in the camel have not been as extensively investigated as in other animal species. The data collected regarding these problems have been derived mainly from information provided by camel owners and from slaughterhouse material. Very limited information has come from clinical and farm observations [5,7,9–11]. Examination of abattoir specimens has provided some information about possible diseases encountered in the reproductive tract, which include pyometra, bursal and ovarian adhesions, endometritis associated with a partially
A. Ali et al. / Theriogenology 84 (2015) 452–456
involuted uterus, and cystic ovarian degeneration [10]. In addition, more recent clinical studies have indicated ovarian hydrobursitis as being an important cause for infertility in dromedary camels [12–15]. Nawito [9] has recorded the bacteriologic findings in the uteri of 2075 dromedary camels of unknown history from Cairo abattoir. In 94 cases, clinical symptoms such as abscesses in the uterus, catarrhal endometritis, hemorrhagic endometritis, pyometra, and pyometra with macerated fetuses were found, along with the presence of Micrococcus pyogenes var. aureus as a prominent factor. Furthermore, the researcher was able to isolate Micrococcus pyogenes var. albus, beta-hemolytic streptococci, Escherichia coli, and Pseudomonas aeruginosa from the uteri of animals that had exhibited clinical symptoms. Studies on the etiology of reproductive disorders in female camels based on a large clinical database are needed to investigate the magnitude of each problem and to plan efficient strategies for their management. The objective of this study was to investigate the clinical findings in large number of barren female dromedaries examined for different complaints. 2. Materials and methods 2.1. Animals and management A total of 7300 female camels (Camelus dromedarius) aged between 5 and 18 years were examined for repeat breeding with regular heat interval (animals that show estrus at regular intervals, 12–14 days, but fail to conceive after three or more services, RB-R, n ¼ 5444), refused mating (a female camel that curls her tail dorsally when approached by a rutting male, RM, n ¼ 1299), repeat breeding with long heat interval (animals which refuse the male 2 weeks after mating as if they were pregnant; however, later on, >30 days, they return to estrus and accept the male, RB-L, n ¼ 489), difficulty or bleeding during mating (DM, n ¼ 53), and for manifestation of malelike behavior including vocalization, chasing estrus females, protrusion of the soft palate, and refusing to mate (MB, n ¼ 15). The average durations (standard deviation) of these reproductive disorders were 5 2.4, 17.5 12, 12 5.6, 7.8 2.5, and 9.5 1.8 months for RB-R, RM, RB-L, DM, and MB groups, respectively. The study was carried out during seven breeding seasons (from September 2007–February 2014). The examinations were conducted at the Veterinary Teaching Hospital of Qassim University, Saudi Arabia. Of the animals examined, 2263 (31%) were nullipara and 5037 (69%) were multipara. Body condition scoring, on a scale of 1 to 5 [16], ranged from 2.5 to 4. All examined females were generally healthy with no systemic illnesses. The majority of animals were left unconfined in open desert areas and fed mainly on alfalfa hay and barley concentrate. They were continuously exposed to fertile males during the breeding season. 2.2. Gynecologic examinations The reproductive tract of each animal was examined through standard transrectal palpation and by
453
ultrasonography using the Aloka SSD-500 model equipped with a 5-MHz linear-array transducer (Aloka Co., Ltd., Tokyo, Japan). The ovaries were examined for structure and size. The uterus was palpated for consistency, movability, and contents. Vaginal examination with a gloved hand was performed to estimate the patency of the vagina and cervix and to evaluate the nature of the vaginal discharges. Catarrhal (turbid mucus), mucopurulent (turbid mucus with flakes of pus), or purulent (profuse pus) vaginal discharges were regarded as signs of clinical endometritis [17–19]. According to the amount and viscidity of the vaginal discharges, degrees of clinical endometritis were categorized as (1) mild: females with small amount of mucopurulent vaginal discharge; (2) moderate: females with moderate mucopurulent discharge; and (3) severe: females with highly viscid and thick or pasty vaginal discharges. 2.3. Bacteriologic examinations A total of 45 female camels with repeat breeding syndrome (failure of conception in spite of mating with a fertile bull more than three times) were randomly selected for bacteriologic examination. Cervical swabs were obtained under aseptic conditions. All cervical samples were inoculated onto blood and MacConkey’s agar plates as described by Collee et al. [20]. The inoculated plates were incubated at 37 C for 48 hours. Gram-stained films were prepared from the growing colonies, and isolates were tested for motility. Gram-negative bacilli and gram-positive cocci were identified by means of API 20E and API 10S systems (Biomerieux SA, Montalien Vercica, France). A commercially available latex agglutination test was used for identification of Lancefield groups of Streptococci. 2.4. Statistical analysis The effect of parity on the frequency of different clinical findings in barren female dromedaries was evaluated using the chi-square test. GraphPad Prism program for Windows, version 300 (1999) was used for analysis. Significance was set at P < 0.05. 3. Results Clinical findings for the different forms of reproductive disorders are shown (Table 1). Clinical endometritis, ovarian hydrobursitis (fluid accumulation and encapsulation of the ovary by the ovarian bursa), and vaginal adhesions (occlusion of the vaginal passage) were the common clinical findings in female camels examined for RB-R, RM, and RB-L, respectively. Vaginal adhesions, persistent hymen, and vulvar atresia were the clinical findings in female camels presented due to bleeding at mating or with a history of an incomplete intromission of the penis (DM). Parity affected the frequency of occurrence of these clinical findings. Clinical endometritis and ovarian hydrobursitis were the common clinical findings in the barren multipara and nullipara, respectively (Table 2). Clinical endometritis was found in mild, moderate, and severe degrees in frequencies of 45%, 32%, and 23%, respectively. Vaginal discharges were observed after
454
A. Ali et al. / Theriogenology 84 (2015) 452–456
Table 1 Clinical findings in female camels examined for repeat breeding with regular heat interval (RB-R, n ¼ 5444), refused mating (RM, n ¼ 1299), repeat breeding with long heat interval (RB-L, n ¼ 489), and difficulty or bleeding at mating (DM, n ¼ 53). Clinical findings
RB-R
Clinical endometritis Ovarian hydrobursitis Vaginal adhesion Cervical stenosis Cervical adhesion Apparently normal genitalia Unovulatory follicles Inactive ovaries Hydrosalpinx Uterine adhesion Pneumovagina Persistent hymen Vulvar atresia Pelvic abscess Total
RM
RB-L
DM
Total
1890 (34.7) 1650 (30.3) 701 (12.9) 269 (4.9) 270 (5) 321 (5.9) 159 (2.9) 50 (0.9) 87 (1.6) 31 (0.6) 16 (0.3) d d
217 (16.7) 140 (10.8) 310 (23.9) 233 (17.9) 214 (16.5) 43 (3.3) 21 (1.6) 85 (6.5) 6 (0.5) 30 (2.3) d d d
152 (31.1) 245 (50.1) 6 (1.2) 28 (5.7) 5 (1) 20 (4.1) d d 33 (6.8) d d d d
5444
1299
489
d d 34 (64.2) d d d d d d d d 12 (22.6) 5 (9.4) 2 (3.8) 53
2259 (31) 2035 (27.9) 1051 (14.4) 530 (7.3) 489 (6.7) 384 (5.3) 180 (2.5) 135 (1.9) 126 (1.7) 61 (0.8) 16 (0.2) 12 (0.2) 5 (0.07) 2 (0.03) 7285
n (%)
exploration of the vagina but rarely during transrectal palpation. Additionally, uterine adhesions to the broad ligaments, rumen, or omentum were recorded. Bilateral and unilateral left and right ovarian hydrobursitis were observed at frequencies of 42%, 46%, and 12%, respectively. The affection was associated with clinical endometritis (21%), adhesions of the genital tract with the internal organs (15%), adhesion of the vagina and pyometra (8%), and distension of the fallopian tube (6%). The other cases had no apparent abnormalities in the genital tract other than the affected bursa. Vaginal adhesions were associated with the accumulation of hypoechogenic or hyperechogenic materials in the uterus in 92% of the cases. No uterine contents were found in the other cases. The vaginal adhesions were found at the caudal-, mid-, or cranial vagina in frequencies of 18%, 74%, and 8%, respectively. A hydrosalpinx could be palpated per rectum as a distended, fluctuated tortuous structure and detected ultrasonographically by the presence of hypoechogenic fluid within the fallopian tube. The affection was mostly unilateral (91.3%). Unovulatory follicles having either thick or thin walls were observed. Those with thick walls and internal trabeculae comprised 92.8%, and they were mostly multiple (79.6%). Persistent hymen was mostly (75%) accompanied by an accumulation of creamy fluids in the anterior vagina. Vulvar atresia characterized by complete occlusion of the vulva lips, with only a very small orifice found at the ventral commissure. One of these cases was accompanied by narrowing of the vagina. The male-like behavior was associated with an enlargement of the clitoris (12 of 15) and narrowing of the vulva and vagina (9 of 15). All these cases were nullipara and aged from 6 to 8 years. Seven cases had normal estrus behavior for about 8 to 12 months before showing masculinity. No ovarian cysts or tumors were detected in any of these cases. A total of 78 isolates were obtained from 39 of the 45 repeat breeding animals randomly selected for bacteriologic investigations; the other six animals had no bacterial isolates. Trueperella pyogenes (formerly named
Arcanobacterium pyogenes) and Staphylococcus aureus were the most commonly found isolates (Table 3). 4. Discussion According to the present data, clinical endometritis, ovarian hydrobursitis, and vaginal adhesions are the main clinical findings in the barren female dromedary camels. Generally, unhygienic measures during parturition, postpartum complications, and unsanitary gynecologic manipulation are contributing factors for uterine infections [7,21]. Venereal infection may also be involved. Pseudomonas aeruginosa, Klebsiella pneumonia, and Streptococcus zooepidemicus, which were reported in this study, are venereal diseases of concern in horses [22,23]. In agreement with the results of this study, Trueperella pyogenes, Staphylococcus aureus, Streptococcus zooepidemicus, Staphylococcus epidermidis, and b-hemolytic Streptococcus have been considered as common causes of endometritis in camels [7,9,24–26]. Endometritis causes
Table 2 Effect of parity on the incidence of different clinical findings in barren female dromedaries (n ¼ 7300). Clinical findings
Nullipara
Clinical endometritis Ovarian hydrobursitis Vaginal adhesion Cervical stenosis Cervical adhesion Apparently normal genitalia Unovulatory follicles Inactive ovaries Hydrosalpinx Uterine adhesion Pneumovagina Male behavior Persistent hymen Vulvar atresia Pelvic abscess Total
636 (28.1) 840 (37.1) 128 (5.7) 25 (1.1) 22 (1) 311 (13.7) 86 (3.8) 102 (4.5) 74 (3.3) 6 (0.3) 0 (0.0) 15 (0.7) 12 (0.5) 5 (0.2) 1 (0.04) 2263
Multipara
P value
1623 (32.2) 1195 (23.7) 923 (18.3) 505 (10) 467 (9.3) 73 (1.5) 94 (1.9) 33 (0.7) 52 (1) 55 (1.1) 16 (0.3) 0 (0.0) 0 (0.0) 0 (0.0) 1 (0.002) 5037
0.004 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.01 0.03 0.02 0.01 0.01 0.08 0.3
n (%)
A. Ali et al. / Theriogenology 84 (2015) 452–456 Table 3 Types and frequencies of cervical bacterial isolates in female dromedary camels presented for repeat breeding syndrome (n ¼ 45 animals). Bacterial isolates
Frequencies ()
Trueperella pyogenes Staphylococcus aureus Streptococcus pyogenes Corynebacterium diphtheroides Bacillus anthracoides Streptococcus zooepidemicus Staphylococcus epidermidis Enterococci Streptococcus b-hemolytica Coagulase –ve Staphylococcus Escherichia coli Proteus Pseudomonas aeruginosa Klebsiella pneumonia Total
14 13 9 8 7 7 5 5 2 2 2 2 1 1 78
infertility for the duration of the infection, and subfertility may persist, even after apparently successful treatment [27]. In addition, early postpartum endometritis has been known to have long-term influences on the steroid concentrations of the ovarian follicles long after the infection self-resolved. This is likely to affect oocyte quality and may partially explain the reduced conception rates and longer interval between calving and conception that are often associated with endometritis [28]. Clinically, most female camels diagnosed with endometritis were repeat breeders, but others refused mating, or accept mating after variable periods of anestrous. Clinical endometritis was detected more frequently in barren multiparous camels. This may be due to the fact that the uterus insulted by a variety of organisms during mating, clinical examination, parturition, and/or postpartum period [7]. In most females, infections are eliminated by natural uterine defense mechanisms. However, in a proportion of the females, these defense mechanisms fail partially or completely, allowing establishment of an infection [7]. Regimes consisted of intrauterine infusion of 100-mL acriflavin 0.1%, 100-mL lotagen 4%, or 300 mg/100-mL gentamicin sulfate accompanied with administration of PGF2a at infusion and hCG at mating have been found equally efficient in treating female camels affected with endometritis. The 90-day nonreturn and calving rates were 64% and 58.9%, 53.1% and 49.3%, and 53.3% and 42.5%, for the three regimes, respectively [29]. Ovarian hydrobursitis is a serious, long-standing, reproductive disorder in dromedary camels characterized by fluid accumulation and encapsulation of the ovary [12–14]. Many observations have suggested that ovarian hydrobursitis is initially an inflammatory process of the genital tract [13,14]. It seems that Chlamydophila abortus is responsible for the spreading of the ovarian hydrobursitis affection in dromedaries [14]. Chlamydophila could be transmitted by all methods of infection including the venereal route [30–32], which may explain the high incidence of infection among the barren nulliparous and multiparous camels. Surgical ablation of the affected bursa in unilaterally affected cases has been recommended because conception rates after these trials have been highly acceptable [12–14]; however, surgical ablation in bilaterally affected cases is not recommended. It is
455
possible to medically treat small-sized ovarian hydrobursitis (
