Subclinical endometritis is one of subscriber factors of Repeat Breeder Syndrome. The purpose of this survey was to measure the consequence of intrauterine cephapirin benzathine disposal after timed unreal insemination ( TAI ) on the construct rate ( CR ) in repetition breeder dairy cattles. All cattles ( n=335 ) had more than three services with no clinically abnormalcies of the generative piece of land were received the combined protocol ; An ear implant incorporating progestagen added the Ovsynch protocol plus 3rd GnRH disposal 7 vitamin D after TAI. Cows in the intervention group ( TRT ; n=160 ) were administered intrauterine cephapirin 12 H after TAI and untreated cattles served as a control group ( CON ; n=175 ) . Synchronism rate ( 98.8 % in TRT, 97.1 % in CON ) and response to 3rd GnRH ( 88.1 % in TRT, 83.9 % in CON ) were similar between groups. There was no consequence of cephapirin on 31 and 62 yearss CR in repetition breeder cattles ( 43.8 % in TRT and 44.0 % in CON ; 39.4 % in TRT and 40.6 % in CON, severally ) . Therefore, the consequences showed that there was no consequence of station AI cephapirin intervention on bettering CR, nevertheless its idea that the combined protocol was used in this survey may be utile for repetition breeder cattles.
Keywords: Repeat breeder, construct rate, endometritis, cephapirin
Repeat breeder syndrome is a major economic loss in dairy industry due to more insemination cost, increased break uping interval and increased culling rates ( Bartlett et al. 1998 ) . Cows that fail to gestate after more than three inseminations with fertile seeds in the absence of noticeable abnormalcies are classified as repetition breeders ( Zemjanis 1980 ) . Besides the causes of the repetition genteelness are multifactorial, it is likely that subclinical endometritis is an of import subscriber to the repetition breeder syndrome of bovine subfertility ( Noakes et al. 2001 ) . Endometritis is one of the most of import causes of unnatural uterine environment and particularly subclinic endometritis consequences with repetition genteelness. The presence of infective bacteriums in the uterus causes redness, histological lesions of the endometrium, distrupted embryo endurance ( Sheldon et al. 2006 ) . Therefore betterment of the intrauterine environment for embryo endurance is the one of different curative methods for repetition genteelness.
Treatment of bacterial endometritis, with intrauterine extract of antibacterial agents and antibiotics, before or after insemination, resulted in changing grades of success ( Veselinovic et al. 1996 ; Shams-Esfandabadi et Al. 2004 ; Kasimanickam et Al. 2005 ; Ahmadi and Dehghan 2007 ) . Cephapirin, a first-generation Mefoxin antibiotic, is active against Gram-positive beings and anaerobiotic bacteriums, and less active against Gram-negative beings. For this ground Cephapirin appeared to be rational antibiotic pick for intrauterine extract ( Mc Dougall 2001 ; LeBlanc et al. 2002 ) . Kasimanickam et Al. ( 2005 ) reported that a individual intervention with cephapirin significantly improved the generative public presentation of cattles with subclinical endometritis. Veselinovic et Al. ( 1996 ) , reported that cephapirin is sufficient when applied one time in instance of clinical endometritis with low grade. Therefore, the purpose of this survey was to measure the consequence of post-AI Cephapirin benzathine ( cephapirin ) disposal on construct rate in repetition breeder dairy cattles.
MATERIAL AND METHODS
Cattles, lodging and direction
This survey was conducted on a commercial dairy herd ( about 800 breastfeeding dairy cattles ) in South Marmara part, Bursa, Turkey. Herd composing was 2/3 Holstein-Friesian and 1/3 Swedish-Red cattles. The wet cattles were housed in free stall installations and grouped harmonizing to their milk production. All cattles were milked three times a twenty-four hours and fed complete assorted rations based on NRC recommendations ( NRC 2001 ) . Average milk production of the herd was 9.880 kilogram ( 305 vitamin D ) per cow. Daily milk output was collected by the ALPRO™ system ( DeLaval, Sweden ) . Average milk production for each cow was recorded for 7 vitamin D before and after AI. All cattles had their organic structure status scored utilizing a 5-point ( 1=thin to 5=fat ) marking system ( Ferguson et al. 1994 ) . All carnal handling and processs were approved by the Lalahan Livestock Central Research Institute Animal Care Committee.
Synchronization protocol and intervention
A sum of 335 cattles ( 283 Holstein-Frisian and 52 Swedish-Red cattles ) were used in this survey. Cows had more than three inseminations with no clinically abnormalcies of the generative piece of land selected as repetition breeder in this survey. All cattles were received the same timed unreal insemination ( TAI ) protocol -the Ovsynch- added progestagen and administered GnRH seven yearss after TAI. Cows, at random phases of the estrous rhythm, received an ear implant incorporating progestagen ( 3 milligram Norgestomet, Crestar, Intervet, Turkey ) and GnRH disposal ( 10 µg Busereline ethanoate, Receptal, Intervet, Turkey ) , on twenty-four hours 0 and PGF2? disposal ( 500 µg Cloprostenol, Estrumate, CEVA-DIF, Turkey ) at the clip of implant remotion, on twenty-four hours 7 and 2nd GnRH injection, on twenty-four hours 9. Then the cattles unnaturally inseminated, at fixed clip, 16-18 H after the 2nd GnRH intervention, utilizing frozen-thawed seeds from bulls of proved birthrate by farm veterinaries. All inseminated cattles received 3rd GnRH disposal ( 10 µg Busereline ethanoate, Receptal ) seven yearss after AI, on twenty-four hours 17.
All inseminated cattles were indiscriminately divided into two groups. The cattles in the intervention Group ( TRT ) were recevied intrauterine 500 milligram of cephapirin benzathine ( Metricure, Intervet, Turkey ) 12 H after TAI ; the cattles in the Control Group ( CON ) were non received any intervention after TAI. The experimental protocol is summarized in Figure 1.
Cows ‘ ovaries were examined on the twenty-four hours of first GnRH and ear implant disposals and 7 yearss subsequently to find ovulation response to first GnRH intervention by ultrasound machine ( Honda HS 2000 equipped with a 7.5 MHz transducer, Honda, Japan ) . Response to first GnRH was characterized by visual aspect of a new principal luteum ( CL ) on ovaries. Maximum follicular size was measured at the clip of AI. Cows were examined seven yearss after TAI to find ovulation by vanishing of dominant follicle at clip of AI and appearence of new CL and besides examined to mensurate follicle size before 3rd GnRH intervention. Response to the 3rd GnRH disposal was detected 7 yearss subsequently 3rd GnRH application ( Figure 1 ) .
Pregnancy diagnosing was performed 31 yearss post-insemination ( Visualization of a fluid-filled uterine horn with embryologic cysts ) by echography. Pregnancy cheque was done station insemination 62 yearss ( Presence of a foetus ) . Pregnancy loss was recorded when the 2nd cheque founded negative. Concept rate was calculated as the figure of cattles diagnosed pregnant, divided by the figure of cattles having AI.
All statistical processs were performed utilizing the computational package of SAS ( Release 9.2, SAS Institute, Cary, NC ) . For statistical analyses, Breed of breastfeeding cattles was coded as 1 ( Holstein-Friesian ) or 2 ( Swedish-Red ) . The ovulatory response to foremost, 2nd and 3rd GnRH disposal was coded as 0 ( No ) or 1 ( Yes ) . Presence of construct on vitamin D 31 and 62 after insemination was coded as 0 ( Not pregnant ) or 1 ( Pregnant ) . Primiparous and multiparous cattles were coded as 1 or 2, severally.
GLM process was performed in order to analyse the followers ; to compare milk production, yearss in milk ( DIM ) , body status mark ( BCS ) , para, figure of services and follicle size at the clip of AI and at the clip of 3rd GnRH between groups ; to find the consequence of synchronism protocol, para and response to foremost and 3rd GnRH on follicle size at the clip of AI. Chi-square analysis utilizing the PROC FREQ process was used to analyse the followers: to compare of ovulatory response to foremost, 2nd and 3rd GnRH and construct rate ( CR ) at 31. and 62. yearss. Logistic process was used to analyse the followers: the consequence of intervention, milk production, DIM, BCS, follicle size at the clip of AI, response to foremost and 3rd GnRH and engender on construct rate.
Average lactation figure of cattles was similar between groups ( 2.1±0.07 in TRT and 2.2±0.06 in CON, severally ) . The average DIM did non differ between TRT and CON groups, 260.3±6.6 and 269.5±6.4, severally. BCS was found to be similar between TRT and CON groups ( 2.92±0.01 and 3.01±0.03, severally ) . But mean milk production was different ( P=0.03 ) between groups ( 31.4±0.6 kg/d in TRT and 29.5±0.6 kg/d in CON ) . Number of services was non different between groups ( 3.9±0.1 in TRT and 4.1±0.09 in CON ) .
Percentage of cattles that ovulated in response to the first GnRH intervention did non differ in TRT ( 67.5 % , 108/160 ) and CON ( 70.9 % , 124/175 ) groups ( Table 1 ) . Synchronism rate ( response to 2nd GnRH ) was non different between groups ( 98.8 % , 158/160 in TRT and 97.1 % , 170/175 in CON ) ( Table 1 ) . Response to the 3rd GnRH was similar between groups ( 89.2 % , 141/158 in TRT and 82.9 % , 141/170 in CON ) ( Table 1 ) .
Maximum follicular size at the clip of AI did non differ between TRT ( 16.07±0.18 millimeter ) and CON ( 16.15±0.18 millimeter ) groups ( Table 2 ) . However, cows that ovulated in response to first GnRH of Ovsynch had produce smaller ( P=0.01 ) follicle size at the clip of AI ( 15.8±0.15 millimeter ) than those of non-responsive ( 16.5±0.23 millimeter ) . Presence of accesory principal CL at the 31 and 62 yearss gestation scrutinies was similar in groups ( Table 1 ) .
Consequence of cephapirin on CR ; at 31 vitamin D was similar between intervention groups with 43.8 % ( 70/160 ) in TRT and 44.0 % ( 77/175 ) in CON. When 62 vitamin D CR were evaluated in groups, there was no differences between TRT ( 39.4 % ; 63/160 ) and CON ( 40.6 % ; 71/175 ) groups ( Table 3 ) . Milk production, DIM, BCS, para, strain, response to foremost, 2nd and 3rd GnRH disposals, follicle size at the clip of AI and station AI cephapirin intervention did non consequence on CR.
Repeat breeder syndrome is a multifactorial status and there is no accurate method for naming the cause of most single instances. Eventhough there are many causative factors ; hormonal instability and uterine infection are considered as of import aetiologic factors for repetition breeder cattles ( Zemjanis 1980 ) . In this survey, the combined protocol ( the Ovsynch added progestagen plus GnRH disposal 7 vitamin D after AI ) was used for extinguishing etiologic hormonal and direction factors every bit much as possible in repetition breeder cattles ; like anovulation, delayed ovulation, Lipo-Lutin lack, estrus sensing mistakes etc.
Recent surveies consequences showed that when merely Ovsynch protocol was applied to normal breastfeeding dairy cattles, ovulatory response to first GnRH of Ovsynch was detected between 45 and 95 % and synchronism rate was determined between 75 and 90 % ( Pursley et al. 1995 ; Galv & A ; atilde ; Os and Santos 2008 ) . Stevenson et Al. ( 2006 ) reported that synchronism rate after Ovsynch+CIDR as 85-95 % . In this survey, foremost ovulatory response and synchronism rate after combined protocol with Ovsynch+progestagen in repetition breeder cattles produced similar consequences with old surveies ( Gumen et al. 2003 ; Bello et Al. 2006 ) . In add-on, earlier surveies indicated that construct rate was 30 to 45 % after Ovsynch in normal dairy cattles ( Pursley et al. 1995 ; Bello et Al. 2006 ) . In this survey, construct rate was found 44.0 % in CON and 43.8 % in TRT groups. This consequences indicate that combined protocol used in this survey improved birthrate in repetition breeder dairy cattles.
When the responses of the groups to this combined protocol evaluated, response to first GnRH, synchronism rate, response to 3rd GnRH and the follicle size at the clip of AI were similar between the intervention and the control groups ( Table 1 and 2 ) . Besides, there were any differences between groups related with covariant factors as DIM, BCS, figure of services and para. Similarity of the groups at responses to the combined protocol and covariant factors, make the rating of the consequence of post-AI cephapirin disposal in repetition breeder cows more clear.
One of the taking factors to reiterate engendering with a high incidence is metritis and different grades of endometritis due to occupying of specific and non-specific infective agents to uterus ( Hart & A ; icirc ; gan et Al. 1972 ) . Its known that subclinical endometritis particularly at unreal insemination might impair the uterine environment and interrupt the nidation and development of the embryo ( Kaufmann et al. 2009 ) . Therefore, rating the efforts of the riddance of endometritis with several intruterine antibiotic extracts at different periods in repetition breeder or normal cattles has been the aim of several surveies ( Veselinovic et al. 1996 ; Kasimanickam et Al. 2005 ) .
Besides post-AI intrauterin antibiotic disposal is good known in the field, we could accomplish merely a few scientific paper evaluated the consequence of post-insemination intrauterin antibiotic disposal on endometritis ( Gupta et al. 1983 ; Ozturkler et Al. 2001 ; Shams-Esfandabadi et Al. 2004 ) . In these limited surveies, similiar to our consequences, some writers did non found any good consequence of intrauterine antibiotic intervention like our survey ( Gupta et al. 1983 ; Shams-Esfandabadi et Al. 2004 ) . Although Ozturkler et Al. ( 2001 ) did non mentioned the consequence of intrauterin antibiotic intervention utilizing entirely, they found that hCG plus intrauterine antibiotic disposals after insemination beneficial to better birthrate in repetition breeder cattles.
Cephapirin, appeared to be rational antibiotic pick for intrauterine extract with activity against Gram-positive beings and anaerobiotic bacteriums, with no witholding period for milk. In this survey, we thought that a individual dosage of post-AI intrauterine cephapirin disposal can better the CR in repetition breeder lactating cattles due to extinguishing subclinic endometritis without any milk waste. Kasimanickam et Al. ( 2005 ) , reported that a individual intervention with cephapirin significantly improved the generative public presentation of cattles with subclinical endometritis. Nevetheless, in the current survey, Cephapirin disposal did non enhanced CR in repetition breeder cattles. Similarly, in the survey of Ahmadi and Dehghan ( 2007 ) resulted with uneffectiveness of cephapirin disposal, they purposed that uterine lavage plus PGF2? disposal, without cephapirin, may be preferred in the intervention of repetition breeder cattles. Besides there are surveies suggested that PGF2? disposal is more ( Steffan et al. 1984 ) or every bit ( Sheldon and Noakes 1998 ; Kasimanickam et Al. 2005 ) . effectual than intrauterine antibiotics for the betterment of generative public presentation in cattles with endometritis.
Our consequences showed that post-AI intrauterine cephapirin disposal did non heighten construct rate in repetition breeder dairy cattles. Additionally, combined synchronism protocol had been used in this survey ; the progestagen added Ovsynch protocol plus GnRH 7 yearss after AI, improved CR stunningly in repetition breeder dairy cattles.
Table 1. Responses to the combined synchronism protocol at repetition breeder dairy cattles.
The Treatment Group
The Control Group
Response to first GnRH of Ovsynch
67.5 % ( 108/160 )
70.9 % ( 124/175 )
Response to 2nd GnRH of Ovsynch
( Synchronization Rate )
98.8 % ( 158/160 )
97.1 % ( 170/175 )
Response to 3rd GnRH ( 7 vitamin D after AI )
89.2 % ( 141/158 )
82.9 % ( 141/170 )
Presence of accoutrement CL
at 31 yearss gestation
38.2 % ( 54/141 )
40.4 % ( 57/141 )
Presence of accoutrement CL
at 62 yearss gestation
29.1 % ( 41/141 )
28.4 % ( 40/141 )
Table 2 Maximum follicle size at the clip of AI and 3rd GnRH after TAI and difference of follicle size in cattles that respond to first GnRH or non in repetition breeder dairy cattles.
The Treatment Group
The Control Group
Follicle size at the clip of AI
Follicle size at clip of
Ovulatory Response ( + )
Ovulatory Response ( – )
Maximal follicle size
15.8±0.1 a millimeter
16.5±0.23 B millimeter
a, B P=0.01
Table 3 Conception rate after combined protocol at 31 and 62 500 post-insemination in repetition breeder dairy cattles.
The Treatment Group
( n=160 )
The Control Group
( n=175 )
Pregnancy rate at 31 vitamin D
43.8 % ( 70/160 )
44.0 % ( 77/175 )
Pregnancy rate at 62 vitamin D
39.4 % ( 63/160 )
40.6 % ( 71/175 )
10.0 % ( 7/70 )
7.8 % ( 6/77 )
Figure 1. Summary of experimental design demoing combined protocol and cephapirin benzathine ( cephapirin ) intervention in repetition breeder cattles. Combined protocol used to extinguish any possible hormonal jobs in repetition breeder cattles. The protocol consist of Ovsynch plus an ear implant ( 3 mg Norgestomet ) for 7 vitamin D between GnRH-PGF2? and GnRH intervention 7 vitamin D after insemination to heighten serum Lipo-Lutin degree by doing accessary CL. Intrauterine cephapirin were used 12 H after insemination in the intervention group ( TRT ) and untreated cattles served as an control ( CON ) .