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Small Ruminant Research 93 (2010) 61–63

Contents lists available at ScienceDirect

Small Ruminant Research

journa l homepage: www.e lsev ier .com/ locate /smal l rumres

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elationship with gamma glutamyl transferase activity andlutaraldehyde coagulation test of serum immunoglobulin Goncentration in newborn goat kids

bru Yalcin ∗, Ethem Mutlu Temizel, Abdullah Yalcin, Ender Carkungozniv. of Uludag, Fac. of Veterinary Med., Turkey

r t i c l e i n f o

rticle history:eceived 16 October 2009eceived in revised form 22 January 2010ccepted 26 March 2010vailable online 24 April 2010

a b s t r a c t

The importance of blood serum immunoglobulin (IgG) concentration, gamma glutamyltransferase (GGT) activity and glutaraldehyde coagulation test (GCT) duration was com-pared to assess passive transfer status in healthy goat kids. Twenty-one Saanen goat kids(0–15 days old) were used. Blood samples were collected from kids presuckling (day 0,baseline), and on the 1st, 4th, 7th and 15th days. Serum IgG concentration, GGT activityand GCT duration were measured on these days. It was observed that serum IgG concentra-

eywords:erum immunoglobulin G (IgG)amma glutamyl transferase (GGT)lutaraldehyde coagulation test (GCT)assive transfer statusoat kids

tion and GGT activity significantly increased on the 1st, 4th and 7th day of the experiment.However serum GCT duration was decreased significantly in kids on the 1st, 4th, 7th and15th days. It was observed that passive transfer status was sufficient when 10% GCT solu-tion clotted serum within 10 and 15 min in up to 7-day-old and 8–15 days old suckling kids,respectively. Thus, GCT as an indicator of GGT activity is useful, practical, and cheaper than

assive t

aanen GGT in assessing p

. Introduction

Goat kids depend on the passive transfer of colostralmmunoglobulin G (IgG) to provide humoral immunityuring the neonatal period.

Passive transfer of colostral IgG is also a critical determi-ant of short-term health and survival for neonatal goatsBarrington and Parish, 2002; Massimini et al., 2007). Goatids with failure of passive transfer (FPT) have an increasedisk of illness and death until at least 6–7 weeks of age.

In neonatal goats <4 days old, an increased risk ofllness and death is associated with serum IgG concentra-ions <1200 mg/dl (O’Brien and Sherman, 1993). Therefore,erum immunoglobulin concentration is an important indi-

∗ Corresponding author at: Department of Internal Medicine, Univ. ofludag, Fac. of Veterinary Med., Animal Teaching Hospital, Campus oforukle, 16059 Bursa, Turkey. Tel.: +90 224 29 40 811;

ax: +90 224 29 40 873.E-mail address: yalcine@uludag.edu.tr (E. Yalcin).

921-4488/$ – see front matter © 2010 Elsevier B.V. All rights reserved.oi:10.1016/j.smallrumres.2010.03.017

ransfer status in goat kids.© 2010 Elsevier B.V. All rights reserved.

cator of FPT (O’Brien and Sherman, 1993; Constant et al.,1994; Arguello et al., 2004; Massimini et al., 2007). Sev-eral methods are available to detect FPT in calves, lambsand kids (Weaver et al., 2000; Maden et al., 2003; Radostitset al., 2007; Lee et al., 2008). Currently, the most accuratemethods of measuring serum IgG concentration are radialimmunodiffusion (RID) and ELISA. However, these meth-ods are impractical for field situations because RID resultsare not available for 18–24 h and ELISA requires labora-tory conditions (Weaver et al., 2000; Massimini et al., 2007;Lee et al., 2008). Another, semiquantitative assay is serumgamma glutamyl transferase (GGT) activity. But a spec-trophotometer or other laboratory apparatus is requiredfor accurate determination of GGT activity (Bostedt et al.,1997; Maden et al., 2003). Glutaraldehyde coagulation test(GCT) which is usually done semiquantitatively, is however

simple, fast, economic and especially available in the fieldconditions (Smith and Sherman, 1994; Batmaz, 1996; Senet al., 2000; Lee et al., 2008).

The aim of this study was to compare the relationbetween GCT duration and GGT enzyme activity, IgG

62 E. Yalcin et al. / Small Ruminant Research 93 (2010) 61–63

Table 1IgG, GGT and GCT findings (mean + SE) in newborn goat kids (n: 21).

Presuckling 1 day 4 days 7 days 15days

IgG (mg/dl) 115.44 ± 6.12bcde** 2423.06 ± 190.6acde** 6722.54 ± 553.5abe** 6661.66 ± 341.7abe** 1330.34 ± 86.8abcd**

GGT (U/l) 87.19 ± 25.6bcd* 743.81 ± 121.23a*cde** 162.08 ± 20.2ad*be** 130.42 ± 21.1ac*be** 51.17 ± 4.23bcd**

** ** * 1 ** ** * **

ifferent

on average 6.5 times greater than pre-suckle levels (Smithand Sherman, 1994). GGT activities reached peak level thesame time in lambs and kids (Bostedt et al., 1997; Britti etal., 2005; Maden et al., 2003) and decreased 48 h after birth.

Table 3Regression analysis between IgG concentrations and GGT activities andGCT durations.

Days Formulas R2

GCT (min.) 60.00 ± 0.0bcde 13.47 ± 2.12ac d

‘a’, ‘b’, ‘c’, ‘d’ and ‘e’ represent differences between the values involving d* p < 0.05.

** p < 0.001.

concentrations in blood serum to assess passive transferstatus in healthy goat kids.

2. Materials and methods

In this study, 21 farmed Saanen goat kids (0–15 days) were used. Thekids were subjected to the same conditions and were not weaned until 1month of age. Blood samples were collected from kids after birth presuck-ling (day 0, baseline), and on the 1st (24 h), 4th, 7th and 15th day of thisstudy. Serum was obtained after centrifugation and stored at −20 C untilanalyzed.

The immunoglobulin concentrations in the serum samples were esti-mated by a quantitative spectrophotometric zinc sulfate turbidity assay(Sherman et al., 1990). The IgG concentrations were derived from a stan-dard curve generated for caprine IgG (Sigma; Cat. No. I5256) (Shermanet al., 1990; Mellado et al., 1998, 2008). GGT activity was assayed usingan enzymatic colorimetric test (Roche Cobas Integra 400 Plus). GCT wasperformed using a 10% glutaraldehyde solution (Merck) (Batmaz, 1996).

Data were analyzed by Friedman Test and Wilcoxon Signed Ranks Test(SPSS for Windows version 13.0) in order to detect statistically signifi-cant differences. Correlations were determined between immunoglobulinconcentrations and GGT activities and GCT duration. Regression analysis(simple and multiple) was performed to evaluate the correlation betweenserum IgG concentration and GGT activity and GCT duration by Pearsoncorrelation test.

3. Results

All kids were clinically examined 1 month after parturi-tion, and they were healthy. Serum IgG concentration, andGGT activity significantly increased on the 1st, 4th and 7thdays of the experiment compared to baseline values. Theresults are summarized in Table 1.

Correlations between IgG concentrations and GGT activ-ities and GCT periods are shown in Table 2. Independentvariables (GGT and GCT) and dependent variable (IgG) werecompared by logistic regression analysis (Table 3).

4. Discussion

In neonatal goats <4 days old, an increased risk ofillness and death is associated with serum IgG concentra-tions <1200 mg/dl (O’Brien and Sherman, 1993). However,Mellado et al. (1998) reported that kids with serum IgGconcentration >800 mg/dl during the first 24 h of life had

Table 2Correlations between IgG concentrations and GGT activities and GCTdurations.

Presuckling 1 day 4 days 7 days 15 days

Ig and GGT −0.450 0.244 −0.043 0.094 0.571*

Ig and GCT − 0.296 −0.110 −0.212 0.351GGT and GCT – 0.076 0.271 −0.202 0.326

* p < 0.05 represents correlation between the different two variables onsame period that are found to be statistically significant.

.69 ± 0.48ab 8.40 ± 3.91a b 12.59 ± 5.53a

letters on the same row that are found to be statistically significant.

a higher survival rate than kids with lower IgG concentra-tions. Goat kids are agammaglobulinic at birth (Constant etal., 1994); however, some researchers (Maden et al., 2003;Britti et al., 2005) reported that newborn goat kids had IgGin their sera, albeit at low levels. In the current study, wedetected some levels of IgG (115.44 ± 6.12 mg/dl) in seraof neonatal goat kids before suckling. Serum IgG concen-trations were measured as 2423.06 ± 190.6 mg/dl at 24 hafter parturition. It reached a peak level of 6722.54 on the4th day after parturition. Maximum serum IgG value wasreported as 7359.0 mg/dl at 18 h and 5270.0 mg/dl at 24 hafter colostrum ingestion in goat kids (Smith and Sherman,1994; Massimini et al., 2007). In general, IgG in goat kids isknown to reach highest concentration in serum 24 h afterbirth. However, Arguello et al. (2004) reported that peakserum IgG concentrations were observed between 24 and60 h. On the other hand, it was found that IgG concentrationremained elevated up to day 7 after birth in calves (Hadornand Blum, 1997).

Serum GGT activity was reported to be an useful param-eter to assess passive transfer status of calves and lambs(Parish et al., 1997; Maden et al., 2003). In this study,GGT activity reached peak level (743.81 ± 121.23) 24 h afterbirth and this level was on average of 8.5 times greaterthan pre-suckle level (81.19 ± 25.6). In agreement with thisfinding, it has been reported that serum GGT levels were

Simple regression0 Ig0 = 174.21 − 0.888 × GGT0 0.201 Ig1 = 1848.02 + 0.835 × (GGT1) 0.051 Ig1 = 2064.95 + 19.930 × (GCT1) 0.084 Ig4 = 6508.35 − 1.498 × (GGT4) 0.0014 Ig4 = 6494.56 − 112.945 × (GCT4) 0.017 Ig7 = 7049.47 − 1.359 × (GGT7) 0.0087 Ig7 = 6991.10 − 21.970 × (GCT7) 0.0415 Ig15 = 564.73 + 16.264 × (GGT15) 0.32*

15 Ig15 = 1266.08 + 10.472 × (GCT15) 0.12

Multiple regression1 Ig1 = 1602.5 + (0.762 × GGT1) + (18.782 × GCT1) 0.134 Ig4 = 6564.6 − (0.510 × GGT4) − (108.812 × GCT4) 0.017 Ig4 = 6564.6 − (0.510 × GGT4) − (108.812 × GCT4) 0.0615 Ig15 = 358.3 + (21.072 × GGT15) + (4.800 × GCT15) 0.42

* p < 0.05 represents results of simple and multiple regression that arefound to be statistically significant.

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Smith, M.C., Sherman, D.M., 1994. Blood, lymph, and immune systems.

E. Yalcin et al. / Small Rum

GT activity decreased 24 h after parturition in the presenttudy. The decrease in serum GGT activity was rapid prob-bly because of its degradation or deactivation (Britti et al.,005). In addition to this, in parallel with GGT results of goatids, in calves that do not have failure of passive transferad serum GGT levels of >200 IU/l, >100 IU/l and >75 IU/l,hen they were 1, 4 and 7 days old, respectively (Parish

t al., 1997). In this study, GGT activity in serum graduallyecreased in postpartum period of kids. Therefore, passiveransfer status of kids may be accepted as sufficient if theyave serum GGT levels of >500 IU/l, >150 IU/l, >100 IU/l and50 IU/, on their 1st, 4th, 7th, and 15th days postpartum,espectively.

Serum GCT duration was also reported to be useful tossess passive transfer status of calves (Batmaz, 1996; Sent al., 2000; Lee et al., 2008,). Using the GCT, serum fromids with IgG levels less than 1000 mg/dl did not clot after0 min incubation with 10% glutaraldehyde solution (Smithnd Sherman, 1994). In this study, GCT duration also indi-ated hypogammaglobulinemia with negative relationshipgG concentrations in goat kids before colostrum intake.hus, the sera of baseline did not clot up to 60 min. GCTuration decreased while serum IgG concentrations andGT activities increased at 24 h after birth. A negativeorrelation was detected between GCT duration and IgGoncentrations on the 4th, 7th and 15th days of experiments shown in Table 1.

In this study, although the relationships between IgGnd GGT, and GCT did not correlate at the statistical level,he results showed parallelism between days of the experi-

ent (Table 1). GGT activities began to decrease after 24 h,ut IgG concentrations continued to remain high up to 7ays. This may be due to the fact that the half-life of IgG is

onger than GGT. Indeed, IgG concentration of calves wasound to remain at high levels during the 7 days after birthHadorn and Blum, 1997). In parallel with low IgG con-entration, GCT solution did not clot up to 60 min beforeeceiving colostrum in goat kids. When IgG concentrationas high, GCT durations (in min) ranged between 1.69 and

3.47 (mean 7.85) min on the 1st, 4th and 7th days of exper-ment. When IgG concentration decreased on the 15thay again, GCT duration was 12.59 min. Sen et al. (2000)eported that serum IgG concentration and GCT durationanged from 2400 to 5800 mg/dl and 1–5 min in calves fromto 10 days old, respectively.

In conclusion, it was observed that passive transfer sta-

us was sufficient when 10% GCT solution clotted within0 and 15 min using samples obtained 7 and 15 daysfter colostrum suckling. Therefore, GCT is useful, practi-al and cheaper than GGT to assess passive transfer statusn neonatal goat kids in field conditions.

esearch 93 (2010) 61–63 63

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