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Abanico veterinario
versão On-line ISSN 2448-6132versão impressa ISSN 2007-428X
Abanico vet vol.8 no.2 Tepic Mai./Ago. 2018
https://doi.org/10.21929/abavet2018.82.2
Original papers
Effect of the disinfection treatment of used intra-vaginal devices on the concentration of progesterone on cows under dry tropical conditions
1Unit Academic of Veterinary Medicine and Zootechnics No.1 - Autonomous University of Guerrero. Mexico.
2Campus of Biological and Agricultural Sciences, Autonomous University of Yucatan. Mexico.
3Faculty of Veterinary Medicine and Zootechnics - University of Veracruz. Mexico.
4College of Postgraduates. Campus Montecillo, Texcoco. Mexico state. Mexico.
The objective of the study was to compare the effect of disinfection/cleaning of CIDR devices on serum progesterone concentrations (P4) in Suizo x Cebú cows, which received during eight days a new intra-vaginal device (CIDR; T1), used device, sterilized by autoclave (T2) or a used device, disinfected with chlorine (T3). A complete randomized design with three replicates per treatment was applied. The used CIDR devices were used previously in cows. Blood samples were collected at 0, 15, 30 min, as well as at 1, 3 and 6 h and then each 12 h until tenth day , with respect to the moment of the insertion of the CIDR. The P4 concentration was determined by radioimmunoassay. Mean concentrations of P4 were similar among treatments (T1, 1.7 ± 0.9 ng/mL; T2, 1.31 ± 0.65 ng/mL; and T3, 1.33 ± 3.19 ng/mL; P<0.05). In conclusion, the process of disinfection/cleaning of the CIDR devices did not affect the availability of progestogen; therefore the CIDR could be used for estrous synchronization.
Keywords: CIDR; Autoclave; progesterone; cows
El objetivo de este estudio fue comparar el efecto de la desinfección/limpieza de los dispositivos CIDR sobre las concentraciones séricas de progesterona (P4) en vacas Suizo x Cebú, a las cuales se les colocó por ocho días un dispositivo intravaginal (CIDR) nuevo (T1), reutilizado esterilizado en autoclave (T2) o reutilizado desinfectado en cloro (T3). Se utilizó un diseño completamente al azar con tres vacas por tratamiento. Los dispositivos CIDR reutilizados habían sido usados en una ocasión en vacas. Se colectaron muestras sanguíneas a los 0, 15, 30 min, 1, 3 y 6 h y posteriormente cada 12 h hasta el día 10, a partir de la inserción del CIDR (hora 0). La concentración de P4 se determinó mediante radioinmunoensayo. Las medias de las concentraciones de P4 fueron similares entre tratamientos (T1, 1.7 ± 0.9 ng/mL; T2, 1.31 ± 0.65 ng/mL; y T3, 1.33 ± 3.19 ng/mL; P>0.05). En conclusión, el proceso de desinfección/limpieza de los dispositivos CIDR no afectó la disponibilidad del progestágeno, por lo que pueden ser reutilizados en el proceso de sincronización del estro.
Palabras clave: CIDR; Autoclave; progesterona; vacas
Introduction
The intra-vaginal devices with progestin have been used for more than four decades with the objective of controlling the estrous cycles of cows and sheep (Lucy et al., 2001, Martínez et al., 2002). The most widely used device in the United States is the controlled internal drug release device (CIDR), for use in livestock (Macmillan and Peterson 1993, Rathbone et al., 1997). The CIDR is a vaginal insert in the form of T that contains 1.9 g (Canada, Mexico, Japan, Australia and New Zealand) or 1.38 g (United States and other countries) of progesterone (P4) in silicone molded on a nylon column (Rathbone et al., 2002; Mapletoft et al., 2003). The residual content of P4 after a 7-day insertion period of CIDR with 1.38 g, has been reported as 0.72 g (Rathbone et al., 2002); therefore, it could be reused.
Although the manufacturer recommends use only once, the reuse of CIDR has been investigated (Martínez et al., 2003; Stevenson et al., 2003; Colazo et al., 2004). Reused CIDR inserts suppress estrus for at least an additional 7 days, in both dairy and meat cattle (Richardson et al., 2002); no differences were observed in pregnancy rates for cattle artificially inseminated at fixed time with new or used CIDR (Colazo et al., 2004).
Different approaches have been used to clean, disinfect or sterilize CIDR devices; however, there are apparently few reports comparing serum progesterone concentrations produced by second-use devices sterilized in an autoclave. Cerri et al. (2005) did not find differences in plasma concentrations of P4 in dairy cows that received a new device (1.38 g of P4), or an insert disinfected in an autoclave and used for 7 days. More scarce are the studies that compare the serum concentrations of P4 produced by CIDR washed and sterilized in an autoclave.
The objective of this study was to evaluate the effect of the disinfection type (autoclave or in chlorine) received by the CIDR to be reused, on the serum progesterone concentration of Suizo x Cebu cows, in the dry tropics of Michoacán, Mexico.
Material and Methods
Location and climate
The research work was carried out in a ranch located in the community of San Lucas, Michoacán, Mexico (18 ° 35'' Latitude North and 100 ° 47'' West Longitude); which is at an altitude of 300 m a.s.l. The annual rainfall is 906 millimetres; with temperatures that oscillate between 20.2 and 37.6 °C (INEGI, 2005). The climate of the region is tropical dry steppe, where there is a critical dry season from February to June.
Animals and handling
Nine Suizo x Cebú cows were used, anestric and lactating, with 60 days postpartum, average weight of 400 + 10 kg and body condition of 4-5 units in the scale from 1 to 9; where 1 corresponded to an emaciated animal and 9 to an obese one (Ayala et al., 1995).
The cows were free grazing in sorghum stubble pastures (Sorghum bicolor), maize stubble (Zea mays) and native grasses (Paspalum spp., Cynodon dactylon, Bouteloa sp., Etc.); as well as shrubs and leguminous plants. The cows were managed in a system with continuous suckling, where the calf remained with the mother until weaning.
The cyclicity of the cows was evaluated prior to the study by means of two palpations at 10-day intervals, considering that a cow was anestric, when on rectal palpation, on the surface of the ovary, the formation of follicles or the presence of a corpus luteum (Salas-Razo et al., 2011); the latter was confirmed by progesterone concentrations lower than 1 ng/ml (Grajales et al., 2006); which indicates the absence of an active corpus luteum.
Application of the CIDR
Once the physiological state of the cows (anestrics) was diagnosed, a CIDR was inserted intra-vaginally for 8 days; three treated groups were formed: 1) cows that received a new CIDR, which was impregnated with 1.9 g of progesterone (CN; n = 3) (Pfizer Laboratory); 2) cows with a previously used CIDR (reused), which after cleaning was sterilized in an autoclave at 121 ºC (CEA, n = 3); and 3) cows with reused CIDR, which was disinfected in chlorine, diluted in water at a rate of 3 ml of chlorine per liter of water (CDC, n = 3). When the CIDR was placed in all the cows, 1 mg of estradiol cypionate (Wittney Laboratory) was applied intramuscularly. Immediately after the withdrawal and 24 h later, 0.25 mg of estradiol cypionate was applied again.
Progesterone analysis
To determine the P4 concentrations, blood samples were taken at 0, 15, 30, 1, 3 and 6 hours and then every 12 hours, until day 10 from the time of insertion of the CIDR (two days after the withdrawal of the CIDR). The samples were obtained by puncturing the coccygeal vein in Vacutainer tubes, in which the blood was allowed to clot for one hour, and then centrifuged at 2500 rpm for 15 min. The serum was stored in aliquots that were stored at -20 ºC until analysis. The measurement of P4 was carried out using a solid phase radioimmunoassay kit. The sensitivity of the test was 0.04 ng/ml and the intra- and interassay variation coefficients 6 and 8.5%
Statistical analysis
A completely randomized design with three cows (repetitions) per treatment was used. All the data were subjected to one-way analysis of variance, for each measurement moment (0.15 30 min, 1, 3, 6 and 12 h, and later every 12 h), using the GLM procedure (SAS, 2009).
Results and Discussion
The majority (75%) of the maximum peaks of P4 occurred between 6 and 12 h post-insertion of the intra-vaginal device (Figure 1). The mean concentrations (± SD) of serum P4 during the 8 days that the remained device, were similar (P> 0.05; for the CN groups (1.7 ± 0.9 ng/ml), CEA (1.31 ± 0.65 ng/ml) and CDC (1.33 ± 3.19 ng/ml). These results were due mainly to differences produced during the first 6 hours, after the insertion of the CIDR (Figures 1 and 2).
Figure 1 Average concentrations of serum progesterone (P4, ng/mL) in Suizo x Cebú cows produced by new CIDR devices (T1 CN), sterilized in autoclave (T2 CEA) and disinfected in chlorine (T3 CDC).
Figure 2 Average serum progesterone concentrations (P4, ng / ml) in Suizo x Cebú cows produced by new devices (T1 CN), sterilized in autoclave (T2 CEA) and disinfected in chlorine (T3 CDC) during the first 12 h post -insertion.
The serum concentrations of P4 obtained from the new CIDR device during 8 days are comparable to the concentrations reported in another study, using the CIDR with 1.38 g (Rathbone et al., 2002). The P4 concentrations obtained with the sterilized CIDR in an autoclave were similar to those obtained by Martínez et al. (2007), who used a reused device that originally contained 1.9 g of P4 in ovariectomized cows. The concentrations of P4 reached their maximum point in the first 6 h after the insertion, maintaining it until approximately 12 hours; however, the same did not happen for the CIDR sterilized with chlorine, which suffered a considerable decrease in the concentration of P4. This decrease was followed by a constant decrease until the elimination of CIDR on day 8. Macmillan et al. (1991) reported that after insertion of a new CIDR (1.9 g of progesterone) into ovariectomized heifers, plasma progesterone concentrations increased to approximately 8.7 ng/ml for 6 h, and then reduced to 6.8 and 2.5 ng/ml on days 1 and 12 post-insertion, respectively. Cerri et al. (2005) comparing plasma concentrations of P4 after the insertion of a new CIDR or one used for 7 days and sterilized in an autoclave, observed that P4 concentrations increased immediately after insertion, reaching its maximum concentration at 90 minutes; following the same pattern during the rest of the insertion period.
Although there was no significant difference between treatments, it is possible that the low concentrations of P4 observed with the disinfected devices in chlorine compared to the devices sterilized in autoclave, were caused by a prolonged exposure to the disinfectant solution. Therefore, it is very likely that steam sterilization of the device in T3 increased the elution rate during the first hours, after insertion compared to T2. Such an effect indicates that the autoclaving process modifies in some way the structure of the implant or the arrangement of the P4 within the insert.
According to McPhee et al. (1983), the plasma progesterone profiles of the intra-vaginal progesterone devices (PRID) reused and sterilized in gas were lower compared to the PRID treated in autoclave; where post-reintegration steady-state plasma concentrations similar to a new insert were observed. The results were attributed to the formation of a large amount of crystalline P4 on the surface of the PRID in an autoclave.
There are structural and functional similarities between the PRID and CIDR devices, since both devices are constructed using micronized progesterone in silicone rubber skin that is molded in a nylon (CIDR), or stainless steel structure (PRID; Rathbone et al., 1998). Due to this similarity, it is possible that the thermal sterilization process used in the present study may have resulted in the same effect observed in autoclaved PRID devices.
Some researchers (Colazo et al., 2004) have used CIDR immersed in a detergent solution based on povidone iodine for 2 hours; followed by washing, rinsing with water, drying in the air and sterilizing with steam in an autoclave equipment; while others have used restricted schemes, only for chemical disinfection (Van Cleeff et al 1992; Martínez et al., 2007) and gas sterilization (Schmitt et al., 1996b). Padula and Macmillan (2006) demonstrated that changes in the vaginal and uterine microflora in the early postpartum of beef cows occurred during 14 days post-insertion of a CIDR. These authors indicate that the microbial cultures in swabs after the withdrawal of the CIDR, produced intense bacterial growth, being the isolated species Pseudomonas aeruginosa and Actinomyces pyogenes.
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Received: June 30, 2017; Accepted: October 23, 2017
Este es un artículo publicado en acceso abierto bajo una licencia Creative Commons
